https://doi.org/10.52973/rcfcv-e34506
Received: 22/08/2024 Accepted: 26/09/2024 Published: 29/12/2024
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Revista Cientรญfica, FCV-LUZ / Vol. XXXIV, rcfcv-e34506
ABSTRACT
Early diagnosis and timely initiation of effective treatment are
critical for the pneumonic pasteurellosis, which is mostly caused
by Mannheimia haemolytica. But recently, strains of M. haemolytica
resistant to antibiotics have begun to emerge, making the treatment
๎„๎ƒฅ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„‰๎„๎„ซ๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎…จ๎ƒ†๎„ท๎„ƒ๎„ด๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎„ซ๎ƒ๎ƒฅ๎„๎„ซ๎ƒ๎ˆ™๎€ƒ๎ƒฐ๎„ด๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎ƒฐ๎„‰๎„จ๎„๎„ซ๎„ด๎‚ญ๎„Š๎„ด๎€ƒ๎„ด๎„๎€ƒ๎ƒฐ๎„Š๎…’๎ƒ๎„ฎ๎„ด๎ƒฐ๎ƒฆ๎‚ญ๎„ด๎ƒ๎€ƒ
the erm (42), msr (E) and mph (E) genes, which are held responsible
๎ƒฅ๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎„ด๎„๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎„ฎ๎ˆ˜๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ท๎„ซ๎„จ๎„๎„ฎ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ซ๎ƒ๎„ฎ๎ƒ๎„Š๎„ด๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ
was aimed to diagnose the presence of M. haemolytica by IHC
method from lung tissue samples of young and adult cattle, and
calves, also to detect erm (42), msr (E) and mph (E) genes, which are
mostly detected in Pasteurella multocida but recently detected in
M. haemolytica๎ˆ™๎€ƒ๎ƒ…๎…™๎€ƒ๎ฑ๎€๎ด๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎ˆ˜๎€ƒ๎๎„Š๎€ƒ๎„จ๎„ซ๎ƒ๎„ฎ๎ƒ๎„Š๎„ด๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎ˆ™๎€ƒ๎„จ๎‚ญ๎„ซ๎‚ญ๎…จ๎„Š๎€ƒ๎ƒ…๎„ƒ๎„๎ƒ†๎„๎„ฎ๎€ƒ
obtained from 100 young and adult cattle, and calves lung samples,
and whose treatment was not responded, were used. Accordingly, the
presence of M. haemolytica was detected by IHC in a total 52 cases of
in 21 (38%) of the calves, 22 (69%) of the young cattle, and 9 (69%) of
the adult cattle. At least one macrolide resistance gene was found by
realโ€“time PCR method in 75% of these positive cases. It was thought
that the percentage frequency of these genes, which cause resistance
by increasing the minimum inhibitory concentrations of macrolide
group antibiotics used in the treatment, by high amounts, makes the
๎„ด๎„ซ๎ƒ๎‚ญ๎„ด๎„‰๎ƒ๎„Š๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎ƒŒ๎ƒฐ๎…จ๎ƒ†๎„ท๎„ƒ๎„ด๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ†๎‚ญ๎„ท๎„ฎ๎ƒ๎„ฎ๎€ƒ๎„ฎ๎ƒ๎„ซ๎ƒฐ๎„๎„ท๎„ฎ๎€ƒ๎ƒ๎ƒ†๎„๎„Š๎„๎„‰๎ƒฐ๎ƒ†๎€ƒ๎„ƒ๎„๎„ฎ๎„ฎ๎ƒ๎„ฎ๎€ƒ
by causing errors in effective antibiotic selection. Recently, strains
of M. haemolytica resistant to antibiotics have begun to emerge,
๎„‰๎‚ญ๎„๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ด๎„ซ๎ƒ๎‚ญ๎„ด๎„‰๎ƒ๎„Š๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„‰๎„๎„ซ๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎…จ๎ƒ†๎„ท๎„ƒ๎„ด๎ˆ˜๎€ƒ๎€น๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„ซ๎ƒ๎‚ญ๎„ฎ๎„๎„Š๎ˆ™๎€ƒ
it is recommended that this type of prevalence studies should be
renewed periodically due to possible resistance development and the
repeated use of antibiotics with resistive shape should be avoided.
Keywords: Mannheimia haemolytica๎ˆ›๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎ˆ›๎€ƒerm (42);
msr (E); mph (E); macrolide; antibiotic resistance; cattle
RESUMEN
๎€ง๎„ƒ๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎„ด๎ƒฐ๎ƒ†๎„๎€ƒ๎„จ๎„ซ๎ƒ๎ƒ†๎„๎…ก๎€ƒ๎…™๎€ƒ๎ƒ๎„ƒ๎€ƒ๎ƒฐ๎„Š๎ƒฐ๎ƒ†๎ƒฐ๎„๎€ƒ๎„๎„จ๎„๎„ซ๎„ด๎„ท๎„Š๎„๎€ƒ๎ƒŒ๎ƒ๎€ƒ๎„ท๎„Š๎€ƒ๎„ด๎„ซ๎‚ญ๎„ด๎‚ญ๎„‰๎ƒฐ๎ƒ๎„Š๎„ด๎„๎€ƒ๎ƒ๎…ฅ๎ƒ†๎‚ญ๎…ก๎€ƒ
son fundamentales para la pasteurelosis neumรณnica, causada
principalmente por Mannheimia haemolytica. Pero recientemente,
han comenzado a surgir cepas de M. haemolytica resistentes a
๎„ƒ๎„๎„ฎ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎„๎„ฎ๎ˆ™๎€ƒ๎„ƒ๎„๎€ƒ๎„ช๎„ท๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎…ฅ๎ƒ†๎„ท๎„ƒ๎„ด๎‚ญ๎€ƒ๎ƒ๎„ƒ๎€ƒ๎„ด๎„ซ๎‚ญ๎„ด๎‚ญ๎„‰๎ƒฐ๎ƒ๎„Š๎„ด๎„๎€ƒ๎ƒŒ๎ƒ๎€ƒ๎„ƒ๎‚ญ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎ƒ†๎ƒฐ๎„๎„Š๎ƒ๎„ฎ๎ˆ˜๎€ƒ
Por lo tanto, es importante investigar los genes erm (42), msr (E)
y mph (E), que se consideran responsables de la resistencia a los
antibiรณticos. El propรณsito del presente estudio tuvo como objetivo
diagnosticar la presencia de M. haemolytica mediante el mรฉtodo
IHC a partir de muestras de tejido pulmonar de bovinos jรณvenes,
๎‚ญ๎ƒŒ๎„ท๎„ƒ๎„ด๎„๎„ฎ๎€ƒ๎…™๎€ƒ๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ซ๎„๎„ฎ๎ˆ™๎€ƒ๎„ด๎‚ญ๎„‰๎ƒ…๎ƒฐ๎ƒ‘๎„Š๎€ƒ๎ƒฐ๎ƒŒ๎ƒ๎„Š๎„ด๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„ซ๎€ƒ๎„ƒ๎„๎„ฎ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒerm (42), msr (E) y
mph (E), que se detectan principalmente en Pasteurella multocida
pero recientemente en M. haemolytica, mediante el mรฉtodo PCR.
๎€ง๎„Š๎€ƒ๎„Š๎„ท๎ƒ๎„ฎ๎„ด๎„ซ๎„๎€ƒ๎ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎ƒฐ๎„๎€ƒ๎„ฎ๎ƒ๎€ƒ๎„ท๎„ด๎ƒฐ๎„ƒ๎ƒฐ๎…ก๎‚ญ๎„ซ๎„๎„Š๎€ƒ๎ƒ…๎„ƒ๎„๎„ช๎„ท๎ƒ๎„ฎ๎€ƒ๎ƒŒ๎ƒ๎€ƒ๎„จ๎‚ญ๎„ซ๎‚ญ๎…ฅ๎„Š๎‚ญ๎€ƒ๎„๎ƒ…๎„ด๎ƒ๎„Š๎ƒฐ๎ƒŒ๎„๎„ฎ๎€ƒ๎ƒŒ๎ƒ๎€ƒ
100 muestras pulmonares de bovinos jรณvenes, adultos y terneros, y
cuyo tratamiento no tuvo respuesta. Por consiguiente, se detectรณ la
presencia de M. haemolytica mediante IHC en un total de 52 casos,
en 21 (38%) de los terneros, 22 (69%) de los bovinos jรณvenes y 9 (69%)
de los bovinos adultos.. Se encontrรณ al menos un gen de resistencia a
macrรณlidos mediante el mรฉtodo de PCR en tiempo real en el 75% de
estos casos positivos. Se pensaba que la frecuencia de estos genes,
que provocan resistencia al aumentar las concentraciones inhibitorias
mรญnimas de antibiรณticos del grupo de los macrรณlidos utilizados en
๎ƒ๎„ƒ๎€ƒ๎„ด๎„ซ๎‚ญ๎„ด๎‚ญ๎„‰๎ƒฐ๎ƒ๎„Š๎„ด๎„๎ˆ™๎€ƒ๎ƒ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒŒ๎‚ญ๎ƒŒ๎ƒ๎„ฎ๎€ƒ๎ƒ๎„ƒ๎ƒ๎…’๎‚ญ๎ƒŒ๎‚ญ๎„ฎ๎ˆ™๎€ƒ๎ƒŒ๎ƒฐ๎…ฅ๎ƒ†๎„ท๎„ƒ๎„ด๎‚ญ๎€ƒ๎ƒ๎„ƒ๎€ƒ๎„ด๎„ซ๎‚ญ๎„ด๎‚ญ๎„‰๎ƒฐ๎ƒ๎„Š๎„ด๎„๎€ƒ๎ƒŒ๎ƒ๎€ƒ
infecciones generando graves pรฉrdidas econรณmicas, al provocar
errores en la selecciรณn efectiva de antibiรณticos. Recientemente,
han comenzado a surgir cepas de M. haemolytica resistentes a los
๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎„๎„ฎ๎ˆ™๎€ƒ๎„ƒ๎„๎€ƒ๎„ช๎„ท๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎…ฅ๎ƒ†๎„ท๎„ƒ๎„ด๎‚ญ๎€ƒ๎ƒ๎„ƒ๎€ƒ๎„ด๎„ซ๎‚ญ๎„ด๎‚ญ๎„‰๎ƒฐ๎ƒ๎„Š๎„ด๎„๎€ƒ๎ƒŒ๎ƒ๎€ƒ๎„ƒ๎‚ญ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎ƒ†๎ƒฐ๎„๎„Š๎ƒ๎„ฎ๎ˆ˜๎€ƒ๎ฑ๎„๎„ซ๎€ƒ
este motivo, se recomienda se renueven periรณdicamente por posible
desarrollo de resistencias y se evite el uso repetido de antibiรณticos
con forma resistiva.
Palabras clave: Mannheimia haemolytica; neumonรญa fibrinosa;
erm (42); msr (E); mph (E); macrรณlido; resistencia
a antibiรณticos; ganado bovino
Determination of Mannheimia haemolytica and its macrolide antibiotic
resistance genes in ๎…ฅbrinous pneumonia of cattle
Determinaciรณn de Mannheimia haemolytica y sus genes de resistencia a
antibiรณticos macrรณlidos en neumonรญa ๎…ฅbrinosa del ganado bovino
Fatih Hatipoglu
1,4
* , Funda Terzi
2
, Mehmet Burak Ates
1
, Asli Balevi
3
, Ozgur Ozdemir
1
, Mustafa Ortatatli
1
1
Selcuk University, Faculty of Veterinary Medicine, Department of Pathology. Konya, Tรผrkiye.
2
Kastamonu University, Faculty of Veterinary Medicine, Department of Pathology. Kastamonu, Tรผrkiye.
3
Selcuk University, Faculty of Veterinary Medicine, Department of Microbiology. Konya, Tรผrkiye.
4
Kyrgyzโ€“Turkish Manas University, Faculty of Veterinary Medicine, Department of Pathology. Bishkek, Kyrgyzistan.
*Corresponding author: fhatip@selcuk.edu.tr
Mannheimia haemolytica resistance genes of cattle / Hatipoglu et al. ______________________________________________________________
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INTRODUCTION
Bovine respiratory disease (BRD) is a leading cause of illness and
death in beef cattle (Bos taurus) worldwide. Pasteurella multocida and
Mannheimia haemolytica, both of which are Gramโ€“negative bacteria
naturally residing in the upper respiratory tract, are the primary
๎ƒ…๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎„จ๎‚ญ๎„ด๎ƒฌ๎„๎ƒฆ๎ƒ๎„Š๎„ฎ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎„๎„Š๎„ฎ๎ƒฐ๎ƒ…๎„ƒ๎ƒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎€œ๎ด๎€ฃ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎„ฎ๎ƒ๎€ƒ๎ƒ…๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎€ƒ๎ƒ†๎‚ญ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„ท๎„ฎ๎ƒ๎€ƒ
mild to severe clinical symptoms, especially when cattle are subjected
to various stressors, including environmental, managemental, or
infectious challenges [1, 2, 3].
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒฅ๎ƒฐ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒ…๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎„ฎ๎„ท๎„จ๎„จ๎„๎„ซ๎„ด๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„๎„ท๎„ด๎ƒ…๎„ซ๎ƒ๎‚ญ๎„๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„๎ƒฅ๎„ด๎ƒ๎„Š๎€ƒ
polymicrobial, that this disease occurs because of the disruption of
the lung defense system and colonization by opportunistic pathogens
๎ƒ…๎…™๎€ƒ๎ƒ†๎„ซ๎„๎„ฎ๎„ฎ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…“๎ƒ๎‚ญ๎„๎ƒ๎„Š๎ƒ๎ƒŒ๎€ƒ๎ƒŒ๎ƒ๎ƒฅ๎ƒ๎„Š๎„ฎ๎ƒ๎€ƒ๎„‰๎ƒ๎ƒ†๎ƒฌ๎‚ญ๎„Š๎ƒฐ๎„ฎ๎„‰๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„‰๎„๎„ฎ๎„ด๎€ƒ๎„จ๎„ซ๎„๎„‰๎ƒฐ๎„Š๎ƒ๎„Š๎„ด๎€ƒ
๎ƒฅ๎ƒ๎‚ญ๎„ด๎„ท๎„ซ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ฎ๎ƒ๎…’๎ƒ๎„ซ๎ƒ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎„ด๎ƒ๎„Š๎„ฎ๎ƒ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎€ƒ๎ƒ๎…˜๎„ท๎ƒŒ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ
into the alveoli. Although direct effects of lipopolysaccharides on
endothelial cells partly cause this condition, macrophageโ€“derived
๎ƒฐ๎„Š๎ƒฅ๎„ƒ๎‚ญ๎„‰๎„‰๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎„‰๎ƒ๎ƒŒ๎ƒฐ๎‚ญ๎„ด๎„๎„ซ๎„ฎ๎€ƒ๎„ฎ๎„ท๎ƒ†๎ƒฌ๎€ƒ๎‚ญ๎„ฎ๎€ƒ๎ฝ๎•๎€น๎ˆฏฮฑ also play an important
๎„ซ๎„๎„ƒ๎ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ท๎„ฎ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎‚ญ๎„๎„ซ๎ƒ๎ƒ…๎ƒฐ๎ƒ†๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎‚ญ๎ƒ๎„ซ๎„๎„จ๎ƒฌ๎ƒฐ๎„ƒ๎ƒฐ๎ƒ†๎€ƒ๎ƒ†๎„ท๎„ƒ๎„ด๎„ท๎„ซ๎ƒ๎„ฎ๎€ƒ๎„ด๎„๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎ƒ๎€ƒ
๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎„จ๎ƒ๎ƒ†๎ƒฐ๎…ฅ๎ƒ†๎€ƒ๎ƒ๎„ด๎ƒฐ๎„๎„ƒ๎„๎ƒฆ๎…™๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎„ท๎„ฎ๎„ท๎‚ญ๎„ƒ๎„ƒ๎…™๎€ƒ๎„ฎ๎„ท๎ƒ†๎ƒ†๎ƒ๎„ฎ๎„ฎ๎ƒฅ๎„ท๎„ƒ๎€ƒ๎„ท๎„Š๎„ƒ๎ƒ๎„ฎ๎„ฎ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎‚ญ๎„Š๎ƒฐ๎„‰๎‚ญ๎„ƒ๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ
treated with antimicrobial drugs. Subacute or chronic cases are
often colonized by opportunistic bacteria, and the primary bacterial
cause may no longer be found. Immunohistochemical (IHC) is used to
diagnose M. haemolytica๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎๎€พ๎€๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎€ƒ๎ƒ†๎‚ญ๎„Š๎€ƒ๎‚ญ๎„ƒ๎„ฎ๎„๎€ƒ๎ƒ…๎ƒ๎€ƒ๎„ท๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ
where fresh tissue is not available, in archival tissues, in cases of
autolysis and antimicrobial treatment. Serum antibody detection
against M. haemolytica๎€ƒ๎„ท๎„ฎ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ƒ๎ƒ๎„ท๎„๎„๎„ด๎„๎…˜๎ƒฐ๎„Š๎€ƒ๎„Š๎ƒ๎„ท๎„ด๎„ซ๎‚ญ๎„ƒ๎ƒฐ๎…ก๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„ด๎ƒ๎„ฎ๎„ด๎ˆ™๎€ƒ๎€ง๎๎๎ท๎€„๎€ƒ
method and bacterial agglutination test are common research tools.
๎€พ๎„๎…“๎ƒ๎…’๎ƒ๎„ซ๎ˆ™๎€ƒ๎„ด๎ƒฌ๎ƒ๎„ฎ๎ƒ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ƒ๎ƒฐ๎„‰๎ƒฐ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„ท๎„ฎ๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎…ฅ๎ƒ๎„ƒ๎ƒŒ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎ƒฐ๎ƒ๎„ฎ๎€ƒ๎ˆซ4].
๎€น๎„๎„ซ๎€ƒ๎…™๎ƒ๎‚ญ๎„ซ๎„ฎ๎ˆ™๎€ƒ๎„‰๎ƒฐ๎„Š๎ƒฐ๎„‰๎‚ญ๎„ƒ๎€ƒ๎ƒฐ๎„Š๎ƒฌ๎ƒฐ๎ƒ…๎ƒฐ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎ƒ†๎„๎„Š๎ƒ†๎ƒ๎„Š๎„ด๎„ซ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎ˆง๎”๎๎€๎ˆจ๎€ƒ๎„๎ƒฅ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ƒฐ๎‚ญ๎„ƒ๎€ƒ
drugs has been measured by tube dilution and disc diffusion (Kirby
Bauer) techniques to determine antibiotic resistance [5]. However,
the inability to isolate the target bacteria from the samples taken
from antibiotic treated or autolytic tissues and the length of the test
periods are considered as limiting factors for the use of these tests [6].
๎ฝ๎„๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒM. haemolytica against 14โ€“, 15โ€“, and
๎‡ ๎‡ฅ๎ˆฏ๎„‰๎ƒ๎„‰๎ƒ…๎ƒ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎„ฎ๎€ƒ๎…’๎ƒฐ๎‚ญ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ฑ๎€๎ด๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎ˆ™๎€ƒ๎„ฎ๎„จ๎ƒ๎ƒ†๎ƒฐ๎…ฅ๎ƒ†๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ
๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒ๎ƒฌ๎‚ญ๎…’๎ƒ๎€ƒ๎ƒ…๎ƒ๎ƒ๎„Š๎€ƒ๎ƒฐ๎ƒŒ๎ƒ๎„Š๎„ด๎ƒฐ๎…ฅ๎ƒ๎ƒŒ๎€ƒ๎ˆซ7]. Whole genome sequencing of the
bacteria has revealed that these resistance phenotypes result from
the combination of at least three distinct macrolide resistance
๎„‰๎ƒ๎ƒ†๎ƒฌ๎‚ญ๎„Š๎ƒฐ๎„ฎ๎„‰๎„ฎ๎ˆ˜๎€ƒ๎๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„ซ๎„ฎ๎„ด๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎ˆ™๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒ๎„ซ๎…™๎„ด๎ƒฌ๎„ซ๎„๎„‰๎…™๎ƒ†๎ƒฐ๎„Š๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ
methyltransferase gene (erm (42)๎ˆจ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎‚ญ๎„Š๎„ด๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎€ƒ
๎ƒ†๎„๎„Š๎ƒฅ๎ƒ๎„ซ๎„ฎ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎”๎๎ท๎€œ๎€ƒ๎ˆง๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎ˆ™๎€ƒ๎„ƒ๎ƒฐ๎„Š๎ƒ†๎„๎„ฎ๎‚ญ๎„‰๎ƒฐ๎ƒŒ๎ƒ๎ˆ™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ฎ๎„ด๎„ซ๎ƒ๎„จ๎„ด๎„๎ƒฆ๎„ซ๎‚ญ๎„‰๎ƒฐ๎„Š๎€ƒ๎€œ๎ˆจ๎€ƒ๎„ด๎…™๎„จ๎ƒ๎€ƒ
I phenotype, with high resistance to lincosamides and low to moderate
resistance to macrolides and streptogramin B. Conformational changes
๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ฑ๎€ƒ๎„ซ๎ƒ๎ƒฆ๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ๎„ซ๎ด๎•๎€„๎€ƒ๎„จ๎„ซ๎ƒ๎…’๎ƒ๎„Š๎„ด๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎ƒ…๎ƒฐ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ๎„ƒ๎ƒฐ๎„‰๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ
inhibitory effect of macrolides on protein synthesis [8๎ˆฌ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎ƒ๎ƒ†๎„๎„Š๎ƒŒ๎€ƒ
group of isolates contains neither an erm gene nor methylation at
nucleotide A2058 and is resistant to macrolides without associated
๎„ƒ๎ƒฐ๎„Š๎ƒ†๎„๎„ฎ๎‚ญ๎„‰๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎ƒฌ๎‚ญ๎„ซ๎ƒ…๎„๎„ซ๎„ฎ๎€ƒ๎„ด๎…“๎„๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ, msr
(E) and mph (E),๎€ƒ๎…“๎ƒฌ๎ƒฐ๎ƒ†๎ƒฌ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„ฎ๎ƒ๎„ช๎„ท๎ƒ๎„Š๎ƒ†๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ๎…˜๎„จ๎„ซ๎ƒ๎„ฎ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎‚ญ๎„‰๎ƒ๎€ƒ
๎„ฎ๎„๎„ท๎„ซ๎ƒ†๎ƒ๎ˆ™๎€ƒ๎ƒ๎„Š๎ƒ†๎„๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎‚ญ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎ƒ๎…ฉ๎„ท๎…˜๎€ƒ๎„จ๎„ท๎„‰๎„จ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎ƒฐ๎„Š๎‚ญ๎ƒ†๎„ด๎ƒฐ๎…’๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ
๎„จ๎ƒฌ๎„๎„ฎ๎„จ๎ƒฌ๎„๎„ด๎„ซ๎‚ญ๎„Š๎„ฎ๎ƒฅ๎ƒ๎„ซ๎‚ญ๎„ฎ๎ƒ๎ˆ™๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎ƒ๎ƒ†๎„ด๎ƒฐ๎…’๎ƒ๎„ƒ๎…™๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ซ๎ƒŒ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎ƒŒ๎ƒฐ๎„ฎ๎„จ๎„ƒ๎‚ญ๎…™๎„ฎ๎€ƒ
high resistance to a wide range of macrolides and carries three
determinants: erm (42), msr (E), and mph (E) [7].
Studies conducted in many countries have shown that resistance to
macrolide group antibiotics has developed in M. haemoltica, suggesting
that this resistance may be regional. In our geography, it has been
observed that many antibiotics, especially macrolide group antibiotics,
are used for respiratory system diseases in cattle, but mostly no
response to treatment is received. Although there have been some
studies on the antibiotic sensitivity and resistance of M. haemolytica
๎ƒฐ๎„Š๎€ƒ๎„๎„ท๎„ซ๎€ƒ๎ƒ†๎„๎„ท๎„Š๎„ด๎„ซ๎…™๎ˆ™๎€ƒ๎„ด๎ƒฌ๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒฌ๎‚ญ๎„ฎ๎€ƒ๎„Š๎„๎„ด๎€ƒ๎ƒ…๎ƒ๎ƒ๎„Š๎€ƒ๎‚ญ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ๎„ฎ๎„จ๎ƒ๎ƒ†๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„ƒ๎„ƒ๎…™๎€ƒ๎ƒฐ๎„Š๎…’๎ƒ๎„ฎ๎„ด๎ƒฐ๎ƒฆ๎‚ญ๎„ด๎ƒฐ๎„Š๎ƒฆ๎€ƒ
macrolide group antibiotics and the resistance genes formed against
๎„ด๎ƒฌ๎ƒ๎„‰๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒฅ๎‚ญ๎ƒ†๎„ด๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎‚ญ๎„Š๎‚ญ๎„‰๎„Š๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎„๎„ซ๎„‰๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒ…๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ
๎‚ญ๎„Š๎ƒฐ๎„‰๎‚ญ๎„ƒ๎€ƒ๎„๎…“๎„Š๎ƒ๎„ซ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ƒ…๎„ซ๎„๎„ท๎ƒฆ๎ƒฌ๎„ด๎€ƒ๎„ด๎„๎€ƒ๎„จ๎‚ญ๎„ด๎ƒฌ๎„๎„ƒ๎„๎ƒฆ๎…™๎€ƒ
department indicates that there is no improvement despite antibiotic
treatment also suggests that antibiotic resistance may develop.
In this study, it was aimed to diagnose the presence of M. haemolytica
๎ƒ…๎…™๎€ƒ๎๎€พ๎€๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„จ๎‚ญ๎„ซ๎‚ญ๎…จ๎„Š๎€ƒ๎ƒ…๎„ƒ๎„๎ƒ†๎„๎„ฎ๎€ƒ๎„จ๎„ซ๎ƒ๎„จ๎‚ญ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ƒ๎„ท๎„Š๎ƒฆ๎€ƒ๎„ด๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎€ƒ๎„ฎ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒ๎„ฎ๎€ƒ
of young and adult cattle, and calves brought to pathology department
and also to identify erm (42), msr (E) and mph (E) genes, which are mostly
detected in P. multocida but recently detected in M. haemolytica, by PCR
method. As a result of this study, it is intended to make an important
contribution to the treatment plan of the disease by both diagnosing
M. haemolytica and determining the macrolide resistance genes from
๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒin cattle.
MATERIALS AND METHODS
Ethical statement
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒ๎„ด๎ƒฌ๎ƒฐ๎ƒ†๎„ฎ๎€ƒ๎ƒ†๎„๎„‰๎„‰๎ƒฐ๎„ด๎„ด๎ƒ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎€น๎‚ญ๎ƒ†๎„ท๎„ƒ๎„ด๎…™๎€ƒ๎„๎ƒฅ๎€ƒ๎‚“๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎‚ญ๎„ซ๎…™๎€ƒ๎”๎ƒ๎ƒŒ๎ƒฐ๎ƒ†๎ƒฐ๎„Š๎ƒ๎ˆ™๎€ƒ
๎€ง๎…˜๎„จ๎ƒ๎„ซ๎ƒฐ๎„‰๎ƒ๎„Š๎„ด๎‚ญ๎„ƒ๎€ƒ๎€„๎„Š๎ƒฐ๎„‰๎‚ญ๎„ƒ๎€ƒ๎ฑ๎„ซ๎„๎ƒŒ๎„ท๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ด๎ƒ๎„ฎ๎ƒ๎‚ญ๎„ซ๎ƒ†๎ƒฌ๎€ƒ๎€๎ƒ๎„Š๎„ด๎ƒ๎„ซ๎ˆ™๎€ƒ๎ท๎ƒ๎„ƒ๎ƒ†๎„ท๎„๎€ƒ
๎‚€๎„Š๎ƒฐ๎…’๎ƒ๎„ซ๎„ฎ๎ƒฐ๎„ด๎…™๎ˆ™๎€ƒ๎Ž๎„๎„Š๎…™๎‚ญ๎ˆ™๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆ™๎€ƒ๎‚ญ๎„จ๎„จ๎„ซ๎„๎…’๎ƒ๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ๎ˆง๎€„๎„จ๎„จ๎„ซ๎„๎…’๎‚ญ๎„ƒ๎€ƒno: 2017/64).
Animals, study area, period and design
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„‰๎‚ญ๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ๎ƒ†๎„๎„Š๎„ฎ๎ƒฐ๎„ฎ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„๎ƒฅ๎€ƒ๎‚ญ๎€ƒ๎„ด๎„๎„ด๎‚ญ๎„ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎‡ ๎‡Ÿ๎‡Ÿ๎€ƒ๎ƒ…๎„๎…’๎ƒฐ๎„Š๎ƒ๎€ƒ๎‚ญ๎„Š๎ƒฐ๎„‰๎‚ญ๎„ƒ๎„ฎ๎€ƒ
(55 calves [less than 6 months old], 32 young cattle [6โ€“24 months old]
and 13 adult cattle [older than 24 months old]) that were brought to
๎„ด๎ƒฌ๎ƒ๎€ƒ๎€ฃ๎ƒ๎„จ๎‚ญ๎„ซ๎„ด๎„‰๎ƒ๎„Š๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎ฑ๎‚ญ๎„ด๎ƒฌ๎„๎„ƒ๎„๎ƒฆ๎…™๎ˆ™๎€ƒ๎€น๎‚ญ๎ƒ†๎„ท๎„ƒ๎„ด๎…™๎€ƒ๎„๎ƒฅ๎€ƒ๎‚“๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎‚ญ๎„ซ๎…™๎€ƒ๎”๎ƒ๎ƒŒ๎ƒฐ๎ƒ†๎ƒฐ๎„Š๎ƒ๎ˆ™๎€ƒ๎ท๎ƒ๎„ƒ๎ƒ†๎„ท๎„๎€ƒ
๎‚€๎„Š๎ƒฐ๎…’๎ƒ๎„ซ๎„ฎ๎ƒฐ๎„ด๎…™๎ˆ™๎€ƒ๎Ž๎„๎„Š๎…™๎‚ญ๎ˆ™๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆ™๎€ƒ๎ƒ…๎ƒ๎„ด๎…“๎ƒ๎ƒ๎„Š๎€ƒ๎‡ก๎‡Ÿ๎‡ ๎‡ก๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‡ก๎‡Ÿ๎‡ ๎‡ฆ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„จ๎„ฎ๎…™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ
๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎„ฎ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎„ด๎ƒฐ๎ƒ†๎€ƒ๎ƒ†๎„ซ๎ƒฐ๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎€ƒ๎„๎ƒฅ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ
๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎ƒ†๎ƒ†๎„๎„ซ๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎ƒŒ๎ƒ๎„ฎ๎ƒ†๎„ซ๎ƒฐ๎ƒ…๎ƒ๎ƒŒ๎€ƒ
in the Results. All animals were routinely necropsied, and tissue
๎„ฎ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„ด๎‚ญ๎„๎ƒ๎„Š๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎ƒฌ๎ƒฐ๎„ฎ๎„ด๎„๎„จ๎‚ญ๎„ด๎ƒฌ๎„๎„ƒ๎„๎ƒฆ๎ƒฐ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎๎€พ๎€๎€ƒ๎ƒ๎…˜๎‚ญ๎„‰๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎‚ญ๎ƒฅ๎„ด๎ƒ๎„ซ๎€ƒ
macroscopic inspection.
Histopathological method
๎๎„ท๎„Š๎ƒฆ๎€ƒ๎„ฎ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒ๎„ฎ๎€ƒ๎„ด๎‚ญ๎„๎ƒ๎„Š๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„จ๎„ฎ๎ƒฐ๎ƒ๎ƒŒ๎€ƒ๎„ซ๎„ท๎„‰๎ƒฐ๎„Š๎‚ญ๎„Š๎„ด๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎…ฅ๎…˜๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ
๎‡ ๎‡Ÿ๎‰ฆ๎€ƒ๎ƒ…๎„ท๎ƒฅ๎ƒฅ๎ƒ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„๎„ซ๎„‰๎‚ญ๎„ƒ๎ƒŒ๎ƒ๎ƒฌ๎…™๎ƒŒ๎ƒ๎€ƒ๎„ฎ๎„๎„ƒ๎„ท๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ๎„‰๎ƒ…๎ƒ๎ƒŒ๎ƒŒ๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„จ๎‚ญ๎„ซ๎‚ญ๎…จ๎„Š๎€ƒ๎‚ญ๎ƒฅ๎„ด๎ƒ๎„ซ๎€ƒ
๎„ซ๎„๎„ท๎„ด๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ด๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎€ƒ๎„จ๎„ซ๎„๎ƒ†๎ƒ๎„ฎ๎„ฎ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„จ๎„ซ๎„๎ƒ†๎ƒ๎ƒŒ๎„ท๎„ซ๎ƒ๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ด๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎€ƒ๎„ฎ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ
sectioned at a thickness of 5 ฮผ๎„‰๎€ƒ๎„๎„Š๎€ƒ๎‚ญ๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎„ด๎„๎„‰๎ƒ๎€ƒ๎ˆง๎๎ƒ๎ƒฐ๎ƒ†๎‚ญ๎€ƒ๎ด๎”๎€ƒ๎‡ก๎‡ ๎‡ก๎‡ค๎ด๎ฝ๎ˆ™๎€ƒ
๎€บ๎ƒ๎„ซ๎„‰๎‚ญ๎„Š๎…™๎ˆจ๎ˆ™๎€ƒ๎„ฎ๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎€พ๎‚ญ๎ƒ๎„‰๎„๎„ด๎„๎…˜๎…™๎„ƒ๎ƒ๎„Š๎ƒ๎‰ช๎€ง๎„๎„ฎ๎ƒฐ๎„Š๎€ƒ๎ˆง๎€พ๎‰ช๎€ง๎ˆจ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ๎…˜๎‚ญ๎„‰๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ
๎„ท๎„Š๎ƒŒ๎ƒ๎„ซ๎€ƒ๎‚ญ๎€ƒ๎ƒ…๎ƒฐ๎„Š๎„๎ƒ†๎„ท๎„ƒ๎‚ญ๎„ซ๎€ƒ๎„ƒ๎ƒฐ๎ƒฆ๎ƒฌ๎„ด๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒ๎€ƒ๎ˆง๎š๎„ƒ๎…™๎„‰๎„จ๎„ท๎„ฎ๎€ƒ๎€œ๎‚™๎‡ค๎‡ ๎ˆ™๎€ƒ๎ฝ๎„๎„๎…™๎„๎ˆ™๎€ƒ๎Œ๎‚ญ๎„จ๎‚ญ๎„Š๎ˆจ๎ˆ˜๎€ƒ
Photographs were taken of the lesions observed during microscopic
๎ƒ๎…˜๎‚ญ๎„‰๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎ˆง๎š๎„ƒ๎…™๎„‰๎„จ๎„ท๎„ฎ๎€ƒ๎€ฃ๎ฑ๎‡ ๎‡ก๎ˆ™๎€ƒ๎ฝ๎„๎„๎…™๎„๎ˆ™๎€ƒ๎Œ๎‚ญ๎„จ๎‚ญ๎„Š๎ˆจ๎ˆ˜
Immunohistochemical method
๎€น๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎๎€พ๎€๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎ˆ™๎€ƒ๎„ฎ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„ด๎‚ญ๎„๎ƒ๎„Š๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎„ท๎„Š๎ƒฆ๎€ƒ๎„ด๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎„ฎ๎€ƒ๎„๎„Š๎€ƒ๎‡ค๎€ƒฮผm
๎„ด๎ƒฌ๎ƒฐ๎ƒ†๎„๎€ƒ๎„ด๎„๎€ƒ๎‚ญ๎ƒŒ๎ƒฌ๎ƒ๎„ฎ๎ƒฐ๎…’๎ƒ๎€ƒ๎„ฎ๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„ฎ๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎ˆง๎ฑ๎ƒ๎„ซ๎„๎…˜๎ƒฐ๎ƒŒ๎‚ญ๎„ฎ๎ƒ๎€ƒ๎€œ๎„ƒ๎„๎ƒ†๎„๎ˆ™๎€ƒ๎ฑ๎„ซ๎„๎„ด๎ƒ๎ƒฐ๎„Š๎€ƒ๎€œ๎„ƒ๎„๎ƒ†๎„๎ˆ™๎€ƒ
๎ฑ๎„๎„ฎ๎„ด๎€ƒ๎ฑ๎„ซ๎ƒฐ๎„‰๎ƒ๎„ซ๎ˆ™๎€ƒ๎ฑ๎„๎„ƒ๎…™๎„‰๎ƒ๎„ซ๎ˆ™๎€ƒ๎€ฃ๎€„๎€œ๎ˆ™๎€ƒ๎€ฃ๎€„๎€œ๎ˆ™๎€ƒ๎€พ๎ƒ๎„‰๎‚ญ๎„ด๎„๎…˜๎…™๎„ƒ๎ƒฐ๎„Š๎ˆจ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‚ญ๎€ƒ๎ƒฅ๎„ท๎„ƒ๎„ƒ๎…™๎€ƒ๎‚ญ๎„ท๎„ด๎„๎„‰๎‚ญ๎„ด๎ƒฐ๎ƒ†๎€ƒ๎๎€พ๎€๎€ƒ
๎ˆง๎๎ƒ๎ƒฐ๎ƒ†๎‚ญ๎€ƒ๎€œ๎„๎„Š๎ƒŒ๎„‰๎‚ญ๎…˜๎ˆจ๎€ƒ๎„๎ƒฐ๎„ด๎€ƒ๎„จ๎„ซ๎„๎ƒ†๎ƒ๎ƒŒ๎„ท๎„ซ๎ƒ๎ˆ˜๎€ƒ๎€ฃ๎ƒ๎„ด๎‚ญ๎ƒฐ๎„ƒ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„จ๎„ซ๎„๎„ด๎„๎ƒ†๎„๎„ƒ๎„ฎ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ
๎ƒ๎‚ญ๎ƒ†๎ƒฌ๎€ƒ๎„จ๎„ซ๎ƒฐ๎„‰๎‚ญ๎„ซ๎…™๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎„๎ƒŒ๎…™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎‚ญ๎„จ๎„จ๎„ƒ๎ƒฐ๎ƒ†๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„ฎ๎ƒฌ๎„๎…“๎„Š๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ฝ๎€„๎€œ๎๎€ง๎‰๎๎ˆ˜๎€ƒ
๎ท๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎„ฎ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎๎€พ๎€๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ๎…˜๎‚ญ๎„‰๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎„ท๎„Š๎ƒŒ๎ƒ๎„ซ๎€ƒ๎‚ญ๎€ƒ๎ƒ…๎ƒฐ๎„Š๎„๎ƒ†๎„ท๎„ƒ๎‚ญ๎„ซ๎€ƒ๎„ƒ๎ƒฐ๎ƒฆ๎ƒฌ๎„ด๎€ƒ
๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒ๎€ƒ๎ˆง๎š๎„ƒ๎…™๎„‰๎„จ๎„ท๎„ฎ๎€ƒ๎€œ๎‚™๎‡ค๎‡ ๎ˆ™๎€ƒ๎ฝ๎„๎„๎…™๎„๎ˆ™๎€ƒ๎Œ๎‚ญ๎„จ๎‚ญ๎„Š๎ˆจ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒ†๎„ซ๎ƒฐ๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎€ƒ๎„๎ƒฅ๎€ƒ๎„จ๎„๎„ฎ๎ƒฐ๎„ด๎ƒฐ๎…’๎ƒ๎€ƒ
IHC staining determined by pathologists.
TABLE I
IHC staining protocols according to primary antibodies
Primary
antibody
Dilution
Antigen
retrieval
Peroxidase
block
Protein
block
Antibody
incubation
Post Primer Polymer DAB Hem*
Mannheimia
haemolytica
antibody
1:1000
HIER1
โ€“ 100ยฐC
โ€“ 30 min
45 min 30 min 60 min 15 min 15 min 5 min 3 min
* Hem: Hematoxylin
FIGURE 1. Agarose gel image of Mannheimia haemolytica con๎™ถrmed by PCR. M:
Marker (100 bp ladder), 1โ€“3: Mannheimia haemolytica strains (227 bp)
Component Volume
Probe 1 ฮผL
Primer Forward 1 ฮผL
Primer Reverse 1 ฮผL
Master 10 ฮผL
H
2
O 2 ฮผL
Sample 5 ฮผL
Total Volume 20 ฮผL
_____________________________________________________________________________Revista Cientifica, FCV-LUZ / Vol. XXXIV, rcfcv-e34423
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Mannheimia haemolytica primary antibody production for IHC
staining
Con๎…ฅrmation of Mannheimia heamolytica isolate
๎€น๎„๎„ซ๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎ƒ๎…˜๎„ด๎„ซ๎‚ญ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎ˆ™๎€ƒ๎‚ญ๎€ƒ๎ƒ†๎„๎„ƒ๎„๎„Š๎…™๎€ƒ๎„๎ƒฅ๎€ƒM. haemolytica isolates was
transferred to brain heart infusion (BHI) liquid medium and incubated
๎‚ญ๎„ด๎€ƒ๎‡ข๎‡ฆ๎‰ฐ๎€๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎‡ ๎€ƒ๎„Š๎ƒฐ๎ƒฆ๎ƒฌ๎„ด๎ˆ˜๎€ƒ๎‚”๎ƒฐ๎…ก๎‚ญ๎„ซ๎ƒŒ๎‰ฎ๎€ƒ๎€บ๎ƒ๎„Š๎„๎„‰๎ƒฐ๎ƒ†๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎ฑ๎„ท๎„ซ๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎Ž๎ƒฐ๎„ด๎€ƒ๎ˆง๎ฑ๎„ซ๎„๎„‰๎ƒ๎ƒฆ๎‚ญ๎ˆ™
Madison๎ˆ™๎€ƒ๎‚”๎ƒฐ๎„ฎ๎ƒ†๎„๎„Š๎„ฎ๎ƒฐ๎„Š๎ˆ™๎€ƒ๎‚€๎ท๎€„๎ˆจ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„ท๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎ƒ๎…˜๎„ด๎„ซ๎‚ญ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎ƒ†๎ƒ†๎„๎„ซ๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ
๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ซ๎„๎„ด๎„๎ƒ†๎„๎„ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„จ๎„๎„ƒ๎…™๎„‰๎ƒ๎„ซ๎‚ญ๎„ฎ๎ƒ๎€ƒ๎ƒ†๎ƒฌ๎‚ญ๎ƒฐ๎„Š๎€ƒ๎„ซ๎ƒ๎‚ญ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎ˆง๎ฑ๎€๎ด๎ˆจ๎€ƒ๎ˆง๎€บ๎ƒ๎„Š๎ƒ๎€ƒ๎€„๎„‰๎„จ๎€ƒ
PCR System 2400, Applied Biosystems, USA) was performed for
๎ƒ†๎„๎„Š๎…ฅ๎„ซ๎„‰๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎ƒฆ๎‚ญ๎ƒฐ๎„Š๎„ฎ๎„ด๎€ƒ๎‡ ๎‡ฅ๎ท๎€ƒ๎„ซ๎ด๎•๎€„๎€ƒ๎„๎ƒฅ๎€ƒ๎„ฎ๎„ท๎„ฎ๎„จ๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒM. haemolytica. PCR
๎„‰๎ƒฐ๎…˜๎€ƒ๎‡ค๎€ƒฮผ๎๎€ƒ๎„๎ƒฅ๎€ƒ๎‡ค๎…˜๎€ƒ๎„‰๎‚ญ๎„ฎ๎„ด๎ƒ๎„ซ๎€ƒ๎„‰๎ƒฐ๎…˜๎€ƒ๎ˆง๎ท๎„๎„ƒ๎ƒฐ๎„ฎ๎€œ๎ƒฐ๎„๎ƒŒ๎…™๎„Š๎ƒ๎ˆ™๎€ƒ๎€ง๎„ฎ๎„ด๎„๎„Š๎ƒฐ๎‚ญ๎ˆจ๎ˆ™๎€ƒ๎‡ค๎‡Ÿ๎€ƒ๎„Š๎ƒฆ๎ˆกฮผ๎
-1
๎€ƒ๎€ฃ๎•๎€„๎ˆ™๎€ƒ๎‡Ÿ๎ˆ˜๎‡ก๎€ƒ
๎„จ๎„‰๎„๎„ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ๎‚ญ๎ƒ†๎ƒฌ๎€ƒ๎„จ๎„ซ๎ƒฐ๎„‰๎ƒ๎„ซ๎ˆš๎€ƒ๎„จ๎„ซ๎ƒฐ๎„‰๎ƒ๎„ซ๎€ƒ๎€น๎‡ค๎€ƒ๎‡ค๎ˆพ๎ˆฏ๎ฝ๎€บ๎€บ๎€บ๎€๎€„๎€„๎ฝ๎€„๎€๎€บ๎€„๎€„๎€๎ฝ๎€„๎€๎ฝ๎€๎€บ๎€บ๎€บ๎ˆฏ๎‡ข๎ˆพ๎€ƒ
๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„จ๎„ซ๎ƒฐ๎„‰๎ƒ๎„ซ๎€ƒ๎ด๎‡ค๎€ƒ๎‡ค๎ˆพ๎ˆฏ๎€๎ฝ๎ฝ๎ฝ๎€„๎€„๎ฝ๎€๎€บ๎ฝ๎€„๎ฝ๎ฝ๎€๎€บ๎€๎€„๎€บ๎ˆฏ๎‡ข๎ˆพ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎‚ญ๎ƒŒ๎ƒŒ๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎€ƒ๎‡ก๎‡ค๎€ƒฮผ๎๎€ƒ๎„๎ƒฅ๎€ƒ
๎„ท๎„ƒ๎„ด๎„ซ๎‚ญ๎„จ๎„ท๎„ซ๎ƒ๎€ƒ๎…“๎‚ญ๎„ด๎ƒ๎„ซ๎ˆ˜๎€ƒ๎€น๎„๎„ซ๎€ƒ๎ฑ๎€๎ด๎ˆ™๎€ƒ๎‚ญ๎€ƒ๎„จ๎„ซ๎„๎„ด๎„๎ƒ†๎„๎„ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎‡จ๎‡ฃ๎‰ฐ๎€๎€ƒ๎‡ฃ๎€ƒ๎„‰๎ƒฐ๎„Š๎€ƒ๎„จ๎„ซ๎ƒ๎ˆฏ๎ƒŒ๎ƒ๎„Š๎‚ญ๎„ด๎„ท๎„ซ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‡ฃ๎‡Ÿ๎€ƒ
๎ƒ†๎…™๎ƒ†๎„ƒ๎ƒ๎„ฎ๎€ƒ๎ˆง๎‡จ๎‡ฃ๎‰ฐ๎€๎€ƒ๎‡ข๎‡Ÿ๎€ƒ๎„ฎ๎ˆ™๎€ƒ๎‡ค๎‡ค๎‰ฐ๎€๎€ƒ๎‡ข๎‡Ÿ๎€ƒ๎„ฎ๎ˆ™๎€ƒ๎‡ฆ๎‡ก๎‰ฐ๎€๎€ƒ๎‡ข๎‡Ÿ๎€ƒ๎„ฎ๎ˆจ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‡ฆ๎‡ก๎‰ฐ๎€๎€ƒ๎‡ ๎‡Ÿ๎€ƒ๎„‰๎ƒฐ๎„Š๎€ƒ๎…ฅ๎„Š๎‚ญ๎„ƒ๎€ƒ๎ƒ๎…˜๎„ด๎ƒ๎„Š๎„ฎ๎ƒฐ๎„๎„Š๎€ƒ
was used. PCR products were run on agarose gel containing 10 ฮผ๎ƒฆ๎ˆก๎„‰๎
-1
ethidium bromide with a 100 bp ladder (Solisbiodyne, Estonia) and a
๎ƒ…๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„๎ƒฅ๎€ƒ๎‡ก๎‡ก๎‡ฆ๎€ƒ๎ƒ…๎„จ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎‚€๎‚“๎€ƒ๎„ƒ๎ƒฐ๎ƒฆ๎ƒฌ๎„ด๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎ƒ†๎„๎„Š๎„ฎ๎ƒฐ๎ƒŒ๎ƒ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎„จ๎„๎„ฎ๎ƒฐ๎„ด๎ƒฐ๎…’๎ƒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ ๎ˆจ๎ˆ˜
Hyperimmune elicitation
One colony of M. haemolytica๎€ƒ๎ˆง๎€œ๎€พ๎๎ˆจ๎€ƒ๎ƒ†๎„๎„Š๎…ฅ๎„ซ๎„‰๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎ฑ๎€๎ด๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎„ท๎„ฎ๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ
๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒฌ๎…™๎„จ๎ƒ๎„ซ๎ƒฐ๎„‰๎„‰๎„ท๎„Š๎ƒฐ๎„ฎ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„ด๎„ซ๎‚ญ๎„Š๎„ฎ๎ƒฅ๎ƒ๎„ซ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎€ƒ๎‡ ๎‡Ÿ๎‡Ÿ๎€ƒ๎„‰๎๎€ƒ๎„๎ƒฅ๎€ƒ๎„ƒ๎ƒฐ๎„ช๎„ท๎ƒฐ๎ƒŒ๎€ƒ
๎„‰๎ƒ๎ƒŒ๎ƒฐ๎„ท๎„‰๎ˆ™๎€ƒ๎ƒฐ๎„Š๎ƒ†๎„ท๎ƒ…๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎ˆง๎”๎‡ค๎‡Ÿ๎‡ฃ๎‡Ÿ๎€ƒ๎€œ๎ฑ๎ˆ™๎€ƒ๎€ง๎„ƒ๎ƒ๎„๎„ด๎„ซ๎„๎„‰๎‚ญ๎ƒฆ๎ˆ™๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆจ๎€ƒ๎‚ญ๎„ด๎€ƒ๎‡ข๎‡ฆ๎‰ฐ๎€๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ
48 h and inactivated with 0.4% formalin. Sterility controls were
๎„จ๎ƒ๎„ซ๎ƒฅ๎„๎„ซ๎„‰๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎ƒฅ๎„ด๎ƒ๎„ซ๎€ƒ๎ƒฐ๎„Š๎‚ญ๎ƒ†๎„ด๎ƒฐ๎…’๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ด๎„ท๎„ซ๎ƒ…๎ƒฐ๎ƒŒ๎ƒฐ๎„ด๎…™๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒ†๎„ท๎„ƒ๎„ด๎„ท๎„ซ๎ƒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎‚ญ๎ƒŒ๎ƒพ๎„ท๎„ฎ๎„ด๎ƒ๎ƒŒ๎€ƒ
๎ƒ…๎…™๎€ƒ๎”๎ƒ†๎€น๎‚ญ๎„ซ๎„ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎€ฃ๎ƒ๎„Š๎„ฎ๎ƒฐ๎„ด๎„๎„‰๎ƒ๎„ด๎ƒ๎„ซ๎€ƒ๎ˆง๎€ฃ๎€ง๎•๎ˆฏ๎‡ ๎ˆ™๎€ƒ๎€œ๎ƒฐ๎„๎„ฎ๎‚ญ๎„Š๎ˆ™๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆจ๎€ƒ๎‚ญ๎ƒ†๎ƒ†๎„๎„ซ๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ด๎„๎€ƒ
๎”๎ƒ†๎€น๎‚ญ๎„ซ๎„ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎„ท๎ƒ…๎ƒ๎€ƒ๎„Š๎„๎ˆš๎‡ฃ๎€ƒ๎„ฎ๎„๎€ƒ๎‚ญ๎„ฎ๎€ƒ๎„ด๎„๎€ƒ๎ƒฌ๎‚ญ๎…’๎ƒ๎€ƒ๎‚ญ๎„จ๎„จ๎„ซ๎„๎…˜๎ƒฐ๎„‰๎‚ญ๎„ด๎ƒ๎„ƒ๎…™๎€ƒ๎‡ ๎ˆ˜๎‡ก๎€ƒ๎‰๎€ƒ๎‡ ๎‡Ÿ
9
bacteria in
๎‡ ๎€ƒ๎„‰๎๎ˆ˜๎€ƒ๎€œ๎„๎„ด๎ƒฌ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒฆ๎ƒ๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎…’๎‚ญ๎ƒ†๎ƒ†๎ƒฐ๎„Š๎ƒ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„จ๎„ซ๎ƒ๎„จ๎‚ญ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒM. haemolytica
for hyperimmune production. Antigen was obtained by not adding
adjuvant to the inactive culture. Candidate vaccine was prepared by
๎ƒฌ๎„๎„‰๎„๎ƒฆ๎ƒ๎„Š๎ƒฐ๎…ก๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎ˆง๎‚€๎„ƒ๎„ด๎„ซ๎‚ญ๎„ด๎„๎„ซ๎„ซ๎‚ญ๎…˜๎ˆ™๎€ƒ๎๎„ฎ๎„๎„ƒ๎‚ญ๎ƒ…๎ˆ™๎€ƒ๎€บ๎ƒ๎„ซ๎„‰๎‚ญ๎„Š๎…™๎ˆจ the inactivated culture and
๎”๎„๎„Š๎„ด๎‚ญ๎„Š๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎๎ท๎€„๎€ƒ๎‡ก๎‡Ÿ๎‡ฅ๎€ƒ๎‚“๎ƒ๎ƒฆ๎‚ญ๎„ด๎‚ญ๎ƒ…๎„ƒ๎ƒ๎€ƒ๎€บ๎„ซ๎‚ญ๎ƒŒ๎ƒ๎€ƒ๎ˆง๎ท๎ƒ๎„จ๎„จ๎ƒฐ๎ƒ†๎ˆ™๎€ƒ๎€น๎„ซ๎‚ญ๎„Š๎ƒ†๎ƒ๎ˆจ๎€ƒ๎‚ญ๎ƒŒ๎ƒพ๎„ท๎…’๎‚ญ๎„Š๎„ด๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ๎„ช๎„ท๎‚ญ๎„ƒ๎€ƒ
proportions and sterility controls were performed (9 ๎€๎€น๎ด๎‰ฌ๎‡ ๎‡ ๎‡ข๎ˆ˜๎‡ข๎‡ข๎ˆจ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ
prepared antigen and candidate vaccine were stored at 4ยฐC until use.
๎ฝ๎„๎€ƒ๎„จ๎„ซ๎„๎ƒŒ๎„ท๎ƒ†๎ƒ๎€ƒ๎ƒฌ๎…™๎„จ๎ƒ๎„ซ๎ƒฐ๎„‰๎„‰๎„ท๎„Š๎ƒ๎€ƒ๎„ฎ๎ƒ๎„ซ๎„ท๎„‰๎ˆ™๎€ƒ๎‡ฃ๎€ƒ๎„ฎ๎…“๎ƒฐ๎„ฎ๎„ฎ๎€ƒ๎‚ญ๎„ƒ๎ƒ…๎ƒฐ๎„Š๎„๎€ƒ๎„‰๎ƒฐ๎ƒ†๎ƒ๎€ƒ๎ˆงMus musculus)
weighing between 16 and 18 g were used. During antigen and vaccine
administration, study mice were monitored for 60 days (d) in terms
of morbidity and mortality. ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ซ๎ƒ๎„จ๎‚ญ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎ƒŒ๎ƒพ๎„ท๎…’๎‚ญ๎„Š๎„ด๎ˆฏ๎ƒฅ๎„ซ๎ƒ๎ƒ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒฆ๎ƒ๎„Š๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ
๎ƒฐ๎„Š๎ƒพ๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„‰๎ƒฐ๎ƒ†๎ƒ๎€ƒ๎ƒฐ๎„Š๎„ด๎„ซ๎‚ญ๎„‰๎„ท๎„ฎ๎ƒ†๎„ท๎„ƒ๎‚ญ๎„ซ๎„ƒ๎…™๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎‡Ÿ๎ˆ˜๎‡ ๎€ƒ๎„‰๎๎€ƒ๎‚ญ๎„ด๎€ƒ๎‡ค๎€ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎„ด๎ƒ๎„ซ๎…’๎‚ญ๎„ƒ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ
๎‡Ÿ๎ˆ˜๎‡ก๎€ƒ๎„‰๎๎€ƒ๎„๎ƒฅ๎€ƒ๎…’๎‚ญ๎ƒ†๎ƒ†๎ƒฐ๎„Š๎ƒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎‚ญ๎ƒŒ๎„‰๎ƒฐ๎„Š๎ƒฐ๎„ฎ๎„ด๎ƒ๎„ซ๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎„ด๎„ซ๎‚ญ๎„‰๎„ท๎„ฎ๎ƒ†๎„ท๎„ƒ๎‚ญ๎„ซ๎„ƒ๎…™๎€ƒ๎„๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎‡ข๎‡Ÿ
th
d of
๎ƒฐ๎„‰๎„‰๎„ท๎„Š๎ƒฐ๎„ฎ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎ˆ™๎€ƒ๎ƒฅ๎„๎„ƒ๎„ƒ๎„๎…“๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎‚ญ๎„ฎ๎„ด๎€ƒ๎ƒฐ๎„Š๎„ด๎„ซ๎‚ญ๎„‰๎„ท๎„ฎ๎ƒ†๎„ท๎„ƒ๎‚ญ๎„ซ๎€ƒ๎ƒฐ๎„Š๎ƒพ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ๎‡Ÿ๎ˆ˜๎‡ก๎€ƒ๎„‰๎๎€ƒ
of vaccine on the 45
th
d and blood was taken from the heart of the
mice on the 60
th
๎€ƒ๎ƒŒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒ…๎„ƒ๎„๎„๎ƒŒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„๎ƒ๎„จ๎„ด๎€ƒ๎‚ญ๎„ด๎€ƒ๎„ซ๎„๎„๎„‰๎€ƒ๎„ด๎ƒ๎„‰๎„จ๎ƒ๎„ซ๎‚ญ๎„ด๎„ท๎„ซ๎ƒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎‡ข๎‡Ÿ๎€ƒ
๎„‰๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎ƒ๎„Š๎€ƒ๎ƒ†๎ƒ๎„Š๎„ด๎„ซ๎ƒฐ๎ƒฅ๎„ท๎ƒฆ๎ƒ๎ƒŒ๎€ƒ๎ˆง๎•๎€น๎ˆฏ๎‡ก๎‡Ÿ๎‡Ÿ๎ˆ™๎€ƒ๎•๎„ผ๎…’๎ƒ๎ˆ™๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆจ๎€ƒ๎‚ญ๎„ด๎€ƒ๎‡ก๎‡ง๎‡Ÿ๎‡Ÿ๎€ƒ๎€บ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎‡ค๎€ƒ๎„‰๎ƒฐ๎„Š๎€ƒ
to separate the serum and the serum was stored at -20ยฐC until use.
Realโ€“time PCR method and primer design
In M. haemolytica, erm (42), msr (E), mph (E) genes were isolated from
๎„จ๎‚ญ๎„ซ๎‚ญ๎…จ๎„Š๎€ƒ๎ƒ…๎„ƒ๎„๎ƒ†๎„๎„ฎ๎€ƒ๎„๎ƒ…๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ซ๎„๎„ท๎„ด๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ด๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎€ƒ๎ƒฅ๎„๎„ƒ๎„ƒ๎„๎…“๎ˆฏ๎„ท๎„จ๎„ฎ๎€ƒ๎‚ญ๎ƒ†๎ƒ†๎„๎„ซ๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ
๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฐ๎„ด๎€ƒ๎„จ๎„ซ๎„๎ƒ†๎ƒ๎ƒŒ๎„ท๎„ซ๎ƒ๎€ƒ๎ˆง๎ณ๎๎€„๎‚ญ๎„‰๎„จ๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎€น๎€น๎ฑ๎€ง๎€ƒ๎ฝ๎ƒฐ๎„ฎ๎„ฎ๎„ท๎ƒ๎€ƒ๎Ž๎ƒฐ๎„ด๎ˆ™๎€ƒ
๎ณ๎ƒฐ๎‚ญ๎ƒฆ๎ƒ๎„Š๎ˆ™๎€ƒ๎€๎‚ญ๎„ด๎€ƒ๎•๎„๎ˆ˜๎ˆฅ๎๎€ฃ๎ˆš๎€ƒ๎‡ค๎‡ฅ๎‡ฃ๎‡Ÿ๎‡ฃ๎ˆจ๎ˆ˜๎€ƒ
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„จ๎„ซ๎ƒ๎ƒฐ๎„Š๎ƒ†๎„ท๎ƒ…๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ด๎ƒ๎‚ญ๎„ƒ๎ˆฏ๎„ด๎ƒฐ๎„‰๎ƒ๎€ƒ๎ฑ๎€๎ด๎€ƒ๎ˆง๎๎ƒฐ๎ƒฆ๎ƒฌ๎„ด๎€๎…™๎ƒ†๎„ƒ๎ƒ๎„ซ๎‰ฎ๎€ƒ๎‡จ๎‡ฅ๎€ƒ
System, Roche, Germany) 96 Cycle device, 95ยฐC by 10 min; 1 cycle,
๎‡จ๎‡ค๎‰ฐ๎€๎€ƒ๎ƒ…๎…™๎€ƒ๎‡ ๎‡ค๎€ƒ๎„ฎ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‡ฅ๎‡ฃ๎‰ฐ๎€๎€ƒ๎ƒ…๎…™๎€ƒ๎‡ฃ๎‡ค๎€ƒ๎„ฎ๎ˆ™๎€ƒ๎‡ฃ๎‡ค๎€ƒ๎ƒ†๎…™๎ƒ†๎„ƒ๎ƒ๎„ฎ๎ˆ™๎€ƒ๎ƒ†๎„๎„๎„ƒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ
๎‡ข๎‡ฆ๎‰ฐ๎€๎€ƒ๎ƒ…๎…™๎€ƒ๎‡ข๎‡Ÿ๎€ƒ๎„ฎ๎ƒ๎ƒ†๎ˆ™๎€ƒ๎‡ ๎€ƒ๎ƒ†๎…™๎ƒ†๎„ƒ๎ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„ท๎„ƒ๎„ด๎„ฎ๎€ƒ๎„๎ƒ…๎„ด๎‚ญ๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ๎…’๎‚ญ๎„ƒ๎„ท๎‚ญ๎„ด๎ƒ๎ƒŒ๎ˆ˜๎€ƒ๎€น๎„๎„ซ๎€ƒ
๎„‰๎„ท๎„ƒ๎„ด๎ƒฐ๎„จ๎„ƒ๎ƒ๎…˜๎€ƒ๎ฑ๎€๎ด๎ˆ™๎€ƒerm (42), mph (E) and msr (E) primers were designed
as previously reported by Rose et al. [9] (๎ฝ๎€„๎€œ๎๎€ง๎€ƒ๎๎๎ˆจ๎ˆ˜
๎ฝ๎„๎€ƒ๎ƒ๎‚ญ๎ƒ†๎ƒฌ๎€ƒ๎…“๎ƒ๎„ƒ๎„ƒ๎€ƒ๎„ท๎„ฎ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎€ฃ๎•๎€„๎€ƒ๎„‰๎‚ญ๎„ฎ๎„ด๎ƒ๎„ซ๎€ƒhydrolysis probes:
TABLE II
Resistance gene sequences
Gen Direction Primary Sequence (5' โ€“ 3')
Fragment
size (bp)
GenBank
accession No
erm (42)
Forward TGCACCATCTTACAAGGAGT
173 HQ888763
Reverse
CATGCCTGTCTTCAAGGTTT
mph (E)
Forward
ATGCCCAGCATATAAATCGC
271 JF769133
Reverse
ATATGGACAAAGATAGCCCG
msr (E)
Forward
TATAGCGACTTTAGCGCCAA
395 JF769133
Reverse GCCGTAGAATATGAGCTGAT
FIGURE 2. Stages of fibrinous pleuropneumonia. A: Active hyperaemia (asterisks), red hepatisation (red arrows), grey
hepatisation (yellow arrows) and expansion of the pleura due to ๎™ถbrin and in๎™ทammatory edema (white arrows), Lung, H&E,
100ร—. B: Active hyperaemia (asterisks), red hepatisation (red arrows), grey hepatisation (yellow arrows) and enlargement of
the interlobular septum due to ๎™ถbrin and in๎™ทammatory edema (white arrows), H&E, Lung, 100ร—. C: Fibrinonecrotic pneumonia,
stages of ๎™ถbrinous pneumonia and necrosis (arrows), Lung, H&E, 200ร—. D: Fibrinous pneumonia, the oat cells with elongated
nuclei (arrows) in the alveolar lumen, Lung, H&E, 400ร—
Mannheimia haemolytica resistance genes of cattle / Hatipoglu et al. ______________________________________________________________
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RESULTS AND DISCUSION
Current knowledge on the frequency and antimicrobial resistance
of bacterial respiratory pathogens is crucial to guide antimicrobial
choice in the control and treatment of BRD. Etiological causes of bovine
๎„ซ๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎„ด๎„ซ๎‚ญ๎ƒ†๎„ด๎€ƒ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„ช๎„ท๎ƒฐ๎„ด๎ƒ๎€ƒ๎„Š๎„ท๎„‰๎ƒ๎„ซ๎„๎„ท๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ†๎„๎„‰๎„จ๎„ƒ๎ƒ๎…˜๎ˆ˜๎€ƒ๎๎„Š๎€ƒ๎ƒฅ๎‚ญ๎ƒ†๎„ด๎ˆ™๎€ƒ
this infection can be successfully treated with many antibiotic groups
including macrolide group. However, the emergence of M.haemolytica
strains resistant to these antibiotics recently causes significant
complications in the control and treatment of the disease [3, 10].
Macroscopic ๎…ฅndings
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒฐ๎ƒ†๎€ƒ๎ƒฅ๎ƒฐ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„ฎ๎ƒ๎ƒ๎„Š๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎„ท๎„Š๎ƒฆ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„ซ๎ƒ๎ƒŒ๎ˆฏ๎ƒ…๎„ซ๎„๎…“๎„Š๎€ƒ
hepatised areas usually scattered in the cranial and sometimes
caudal lobes, enlargement of the interlobular septum, whiteโ€“yellow
๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎ƒ๎…˜๎„ท๎ƒŒ๎‚ญ๎„ด๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ƒ๎ƒ๎„ท๎„ซ๎‚ญ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎„ซ๎ƒ๎‚ญ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎‚ญ๎ƒŒ๎ƒฌ๎ƒ๎„ฎ๎ƒฐ๎„๎„Š๎€ƒ๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ด๎ƒฌ๎„๎„ซ๎‚ญ๎…˜๎ˆ˜๎€ƒ
๎๎„Š๎€ƒ๎‚ญ๎ƒŒ๎ƒŒ๎ƒฐ๎„ด๎ƒฐ๎„๎„Š๎ˆ™๎€ƒ๎‚ญ๎€ƒ๎„จ๎„ท๎„ซ๎„ท๎„ƒ๎ƒ๎„Š๎„ด๎€ƒ๎ƒ†๎„๎„Š๎„ด๎ƒ๎„Š๎„ด๎€ƒ๎ˆง๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„จ๎„ท๎„ซ๎„ท๎„ƒ๎ƒ๎„Š๎„ด๎€ƒ๎ƒ…๎„ซ๎„๎„Š๎ƒ†๎ƒฌ๎„๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎ˆจ๎€ƒ
๎„๎„ซ๎€ƒ๎‚ญ๎„ซ๎ƒ๎‚ญ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎…’๎‚ญ๎„ซ๎…™๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎„ฎ๎ƒฐ๎…ก๎ƒ๎€ƒ๎ˆง๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎ˆจ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ
sometimes detected on the cut surface.
Histopathologic ๎…ฅndings
Microscopically, an initial period characterized by active hyperaemia
of the capillaries in the interalveolar septum and alveolar edema and
the period of red hepatisation with neutrophil granulocytes, shed
๎ƒ๎„จ๎ƒฐ๎„ด๎ƒฌ๎ƒ๎„ƒ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎ƒ†๎ƒ๎„ƒ๎„ƒ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒŒ๎ƒ๎„Š๎„ฎ๎ƒ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎„ซ๎ƒ๎‚ญ๎ƒŒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎‚ญ๎„ƒ๎…’๎ƒ๎„๎„ƒ๎ƒฐ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎ˆ˜๎€ƒ
Regions belonging to grey hepatisation periods with disappearance
of hyperaemia but intraalveolar neutrophil granulocytes, shed
๎ƒ๎„จ๎ƒฐ๎„ด๎ƒฌ๎ƒ๎„ƒ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎ƒ†๎ƒ๎„ƒ๎„ƒ๎„ฎ๎ˆ™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎ƒ…๎„ท๎„Š๎ƒŒ๎‚ญ๎„Š๎„ด๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎‚ญ๎„ƒ๎„ฎ๎„๎€ƒ๎„Š๎„๎„ด๎ƒ๎ƒŒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ก๎ˆฏ๎€„๎ˆจ๎ˆ˜๎€ƒ๎๎„Š๎€ƒ
addition, enlargement of the interlobular septum was observed due
๎„ด๎„๎€ƒ๎ƒฐ๎„Š๎…ฆ๎‚ญ๎„‰๎„‰๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎ƒ๎…˜๎„ท๎ƒŒ๎‚ญ๎„ด๎ƒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ก๎ˆฏ๎€œ๎ˆจ๎ˆ˜๎€ƒ๎•๎ƒ๎ƒ†๎„ซ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎‚ญ๎„ซ๎ƒ๎‚ญ๎„ฎ๎€ƒ
๎„ฎ๎„ท๎„ซ๎„ซ๎„๎„ท๎„Š๎ƒŒ๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎‚ญ๎„Š๎€ƒ๎ƒฐ๎„Š๎…ฆ๎‚ญ๎„‰๎„‰๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎…ก๎„๎„Š๎ƒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ก๎ˆฏ๎€๎ˆจ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ˆต๎„๎‚ญ๎„ด๎€ƒ๎ƒ†๎ƒ๎„ƒ๎„ƒ๎„ฎ๎ˆถ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ
๎ƒ๎„ƒ๎„๎„Š๎ƒฆ๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„Š๎„ท๎ƒ†๎„ƒ๎ƒ๎ƒฐ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎„๎ƒ†๎ƒ†๎‚ญ๎„ฎ๎ƒฐ๎„๎„Š๎‚ญ๎„ƒ๎„ƒ๎…™๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ก๎ˆฏ๎€ฃ๎ˆจ๎ˆ˜๎€ƒ
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒฐ๎ƒ†๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒฐ๎ƒ†๎€ƒ๎…ฅ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„จ๎„ซ๎ƒ๎„ฎ๎ƒ๎„Š๎„ด๎€ƒ
๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎ƒ†๎„๎„Š๎„ฎ๎ƒฐ๎„ฎ๎„ด๎ƒ๎„Š๎„ด๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎ˆ™๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ
FIGURE 4. A: Antiโ€“Mannheimia haemolytica positive immunoreactivity in in๎™ทammatory cells, desquamated epithelial cells
and alveolar macrophages in alveolar lumens, Lung, IHC, bar: 100 ยตm. B: Anti
โ€“Mannheimia haemolytica positive reaction in
desquamated epithelium and in๎™ทammatory cells in alveolar lumens, IHC, Lung, bar: 50 ยตm. C: Antiโ€“Mannheimia haemolytica
positive reaction in in๎™ทammatory cells, IHC, bar: 50 ยตm. D: IHC, bar: 100 ยตm
Calf Young
cattle
Adult
cattle
0
10
20
30
40
Cases
IHC (-)
IHC (+)
FIGURE 3. Graphical representation of Mannheimia haemolytica positive or
negative cases by IHC
_____________________________________________________________________________Revista Cientifica, FCV-LUZ / Vol. XXXIV, rcfcv-e34423
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๎„๎„ซ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„จ๎„ท๎„ซ๎„ท๎„ƒ๎ƒ๎„Š๎„ด๎€ƒ๎ƒ…๎„ซ๎„๎„Š๎ƒ†๎ƒฌ๎„๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎„จ๎„ซ๎ƒ๎…’๎ƒฐ๎„๎„ท๎„ฎ๎„ƒ๎…™๎€ƒ๎„ซ๎ƒ๎„จ๎„๎„ซ๎„ด๎ƒ๎ƒŒ๎€ƒ๎ˆซ2, 4, 11].
Dorso et al. [2] ๎„ซ๎ƒ๎„จ๎„๎„ซ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„‰๎ƒถ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒถ๎ƒ†๎€ƒ๎ƒฅ๎ƒถ๎„Š๎ƒŒ๎ƒถ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฅ๎ƒถ๎ƒ…๎„ซ๎ƒถ๎„Š๎„๎„ท๎„ฎ๎€ƒ
๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒถ๎‚ญ๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎ƒ†๎ƒฌ๎‚ญ๎„ซ๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒถ๎…ก๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎ƒฅ๎ƒถ๎ƒ…๎„ซ๎ƒถ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„ƒ๎ƒ๎„ท๎„ซ๎ƒถ๎„ด๎ƒถ๎„ฎ๎ˆ™๎€ƒ๎„ƒ๎„ท๎„‰๎ƒถ๎„Š๎‚ญ๎„ƒ๎€ƒ๎ƒ๎…˜๎„ท๎ƒŒ๎‚ญ๎„ด๎ƒ๎€ƒ
๎ƒ†๎„๎„‰๎„จ๎„๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„Š๎ƒ๎„ท๎„ด๎„ซ๎„๎„จ๎ƒฌ๎ƒถ๎„ƒ๎„ฎ๎ˆ™๎€ƒ๎ƒฅ๎ƒถ๎ƒ…๎„ซ๎ƒถ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎‚ญ๎„ƒ๎…’๎ƒ๎„๎„ƒ๎ƒถ๎„ด๎ƒถ๎„ฎ๎ˆ™๎€ƒ๎ˆต๎„๎‚ญ๎„ด๎€ƒ๎ƒ†๎ƒ๎„ƒ๎„ƒ๎„ฎ๎ˆถ๎ˆ™๎€ƒ๎„Š๎ƒ๎ƒ†๎„ซ๎„๎„ด๎ƒถ๎ƒ†๎€ƒ
๎ƒฅ๎„๎ƒ†๎ƒถ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ…๎„ƒ๎„๎„๎ƒŒ๎€ƒ๎…’๎ƒ๎„ฎ๎„ฎ๎ƒ๎„ƒ๎€ƒ๎„๎„ซ๎€ƒ๎„ƒ๎…™๎„‰๎„จ๎ƒฌ๎‚ญ๎„ด๎ƒถ๎ƒ†๎€ƒ๎„ด๎ƒฌ๎„ซ๎„๎„‰๎ƒ…๎„๎„ฎ๎ƒถ๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒ๎‚ญ๎„ซ๎ƒ†๎ƒฌ๎ƒ๎„ซ๎„ฎ๎€ƒ๎‚ญ๎„ƒ๎„ฎ๎„๎€ƒ
recorded that the most frequently detected pathogens among the 26
๎„ƒ๎„ท๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„ฎ๎‚ญ๎„‰๎„จ๎„ƒ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎ƒฌ๎ƒถ๎„ฎ๎„ด๎„๎„ƒ๎„๎ƒฆ๎ƒถ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ฑ๎€๎ด๎€ƒ๎ƒ๎…˜๎‚ญ๎„‰๎ƒถ๎„Š๎‚ญ๎„ด๎ƒถ๎„๎„Š๎„ฎ๎€ƒ๎…“๎ƒถ๎„ด๎ƒฌ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ฎ๎ƒ†๎„๎„จ๎ƒถ๎ƒ†๎€ƒ
๎„ƒ๎ƒ๎„ฎ๎ƒถ๎„๎„Š๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎”๎ˆ˜๎€ƒ๎ƒฌ๎‚ญ๎ƒ๎„‰๎„๎„ƒ๎…™๎„ด๎ƒถ๎ƒ†๎‚ญ (69.2%) and ๎ฑ๎ˆ˜๎€ƒ๎„‰๎„ท๎„ƒ๎„ด๎„๎ƒ†๎ƒถ๎ƒŒ๎‚ญ (61.5%).
Immunohistochemical ๎…ฅndings
Accordingly, the presence of M. haemolytica was detected by IHC
in 21 of 55 calves (38%), 22 of 32 young cattle (69%) and 9 of 13 adult
๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎€ƒ๎ˆง๎‡ฅ๎‡จ๎‰ฆ๎ˆจ๎ˆ™๎€ƒ๎„ด๎„๎„ด๎‚ญ๎„ƒ๎„ƒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎‡ค๎‡ก๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ˆง๎‡ค๎‡ก๎‰ฆ๎ˆจ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ข๎ˆจ๎ˆ˜๎€ƒ
Immunoreactivity was found in the lumens of alveoli, bronchi and
bronchioles, in the cytoplasm of neutrophil granulocytes and alveolar
๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„จ๎ƒฌ๎‚ญ๎ƒฆ๎ƒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„ซ๎ƒ๎‚ญ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„Š๎…ฆ๎‚ญ๎„‰๎„‰๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ฎ๎„๎„‰๎ƒ๎„ด๎ƒฐ๎„‰๎ƒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ…๎„ซ๎„๎„Š๎ƒ†๎ƒฌ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ
๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒ…๎„ซ๎„๎„Š๎ƒ†๎ƒฌ๎ƒฐ๎„๎„ƒ๎‚ญ๎„ซ๎€ƒ๎ƒ๎„จ๎ƒฐ๎„ด๎ƒฌ๎ƒ๎„ƒ๎ƒฐ๎„ท๎„‰๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ4 Aโ€“Bโ€“Cโ€“D)
In pneumonia cases in cattle in our country; Haziroglu et al. [12]
reported that M .haemolytica was isolated in 42%, P. multocida in
8%, Histophilus somni in 10%, both M. haemolytica and H. somni in
7%, and M. haemolytica and P. multocida in 2% of 100 calf lungs with
pneumonia. Kilic and Muz [13] detected pneumonia in 500 (6%) of
a total of 8,222 cattle lungs slaughtered in an abattoir and isolated
P. multocida in 30 (6%) and M. haemolytica in 9 (1.8%) of them. As
can be understood from these studies, M. haemolytica is one of the
most important bacterial agents in bovine pneumonia in our country.
๎€พ๎„๎…“๎ƒ๎…’๎ƒ๎„ซ๎ˆ™๎€ƒ๎ƒฐ๎„ด๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„Š๎„๎„ด๎€ƒ๎„๎„Š๎„ƒ๎…™๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎„ฎ๎€ƒ๎ƒ…๎„ท๎„ด๎€ƒ
also cases with pneumonia in general and even healthy lungs were
included in a study [14]. In addition, it is inevitable that there may
๎ƒ…๎ƒ๎€ƒ๎ƒŒ๎ƒ๎…ฅ๎ƒ†๎ƒฐ๎ƒ๎„Š๎ƒ†๎ƒฐ๎ƒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎„ซ๎ƒ๎‚ญ๎„ด๎„‰๎ƒ๎„Š๎„ด๎€ƒ๎ƒŒ๎„ท๎ƒ๎€ƒ๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒฐ๎„Š๎‚ญ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎…™๎€ƒ๎„ด๎„๎€ƒ
TABLE III
Distribution of resistance genes according to cases
Calf
(< 6 month)
(n = 55)
Young cattle
(> 6 month โ€“ < 2 year)
(n = 32)
Adult cattle
(> 2 year)
(n = 13)
Total
(n = 100)
IHC (+) 21 (38%) 22 (69%) 9 (69%) 52
erm (42) โ€“ โ€“ โ€“ โ€“
msr (E) 4 2 2 8 (15%)
mph (E) 1 โ€“ 1 2 (4%)
msr (E) + mph (E) 7 7 3 17 (33%)
msr (E) + mph (E) + erm (42) 5 5 2 12 (23%)
No expression 4 8 1 13 (25%)
Accordingly, at least one resistance gene was found in 75% of these positive cases
Mannheimia haemolytica resistance genes of cattle / Hatipoglu et al. ______________________________________________________________
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produce M. haemolytica in bacterial cultures both in these studies
and in laboratory analyses, especially in cases where antibiotics are
used. As a matter of fact, in present study in which only 100 different
๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎ƒ…๎„ซ๎„๎„Š๎ƒ†๎ƒฌ๎„๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ๎…’๎‚ญ๎„ƒ๎„ท๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎…“๎ƒฌ๎ƒฐ๎ƒ†๎ƒฌ๎€ƒ๎ƒŒ๎ƒฐ๎ƒฅ๎ƒฅ๎ƒ๎„ซ๎ƒ๎ƒŒ๎€ƒ
from other studies in this respect, M. haemolytica was found positive
in 52 cases by IHC method. When the distribution of these cases
according to age groups was analysed, the rate of the causative agent,
which was around 38% in calves, could reach up to 69% in young cattle
๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎ƒŒ๎„ท๎„ƒ๎„ด๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ข๎ˆจ๎ˆ˜๎€ƒ๎๎„ด๎€ƒ๎ƒ†๎‚ญ๎„Š๎€ƒ๎ƒ…๎ƒ๎€ƒ๎„ด๎ƒฌ๎„๎„ท๎ƒฆ๎ƒฌ๎„ด๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„ฎ๎ƒฐ๎„ด๎„ท๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„‰๎‚ญ๎…™๎€ƒ๎ƒ…๎ƒ๎€ƒ
because this agent is less common in calves and that calves are not
๎ƒ๎…˜๎„จ๎„๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎€ƒ๎„จ๎„ซ๎ƒ๎ƒŒ๎ƒฐ๎„ฎ๎„จ๎„๎„ฎ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎ƒฅ๎‚ญ๎ƒ†๎„ด๎„๎„ซ๎„ฎ๎€ƒ๎„ฎ๎„ท๎ƒ†๎ƒฌ๎€ƒ๎‚ญ๎„ฎ๎€ƒ๎„ด๎„ซ๎‚ญ๎„Š๎„ฎ๎„จ๎„๎„ซ๎„ด๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎ˆ™๎€ƒ๎ƒ†๎„ซ๎‚ญ๎„‰๎„จ๎ƒ๎ƒŒ๎€ƒ
housing, inadequate nutrition, etc.
On the other hand, the detection of the presence of the agent in
๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒ†๎…™๎„ด๎„๎„จ๎„ƒ๎‚ญ๎„ฎ๎„‰๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ๎„จ๎ƒฐ๎„ด๎ƒฌ๎ƒ๎„ƒ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎…ฆ๎‚ญ๎„‰๎„‰๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎ƒ†๎ƒ๎„ƒ๎„ƒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎ƒ๎„ฎ๎ƒฐ๎„๎„Š๎ƒ๎ƒŒ๎€ƒ
๎‚ญ๎„ซ๎ƒ๎‚ญ๎„ฎ๎€ƒ๎ˆง๎€น๎๎€บ๎€ƒ๎‡ฃ๎ˆจ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‚ญ๎ƒฆ๎„ซ๎ƒ๎ƒ๎„‰๎ƒ๎„Š๎„ด๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„Š๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„๎„ด๎ƒฌ๎ƒ๎„ซ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎ƒฐ๎ƒ๎„ฎ๎€ƒ๎ˆซ15,
16]. In addition, by revealing the presence of the causative agent
with IHC, disadvantages such as antibiotic use, autolysis, and failure
to isolate the causative agent in culture were prevented and more
reliable results were obtained.
Realโ€“Time PCR and resistance genes ๎…ฅndings
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎‚ญ๎ƒฐ๎„ƒ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎€ƒ๎ƒฐ๎„Š๎…’๎ƒ๎„ฎ๎„ด๎ƒฐ๎ƒฆ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„จ๎ƒ๎„ซ๎ƒฅ๎„๎„ซ๎„‰๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ
๎„‰๎„ท๎„ƒ๎„ด๎ƒฐ๎„จ๎„ƒ๎ƒ๎…˜๎€ƒ๎„ซ๎ƒ๎‚ญ๎„ƒ๎€ƒ๎„ด๎ƒฐ๎„‰๎ƒ๎€ƒ๎ฑ๎€๎ด๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎€ƒ๎„๎„Š๎€ƒM.haemolytica positive cases (52)
๎ƒ…๎…™๎€ƒ๎๎€พ๎€๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„ฎ๎„ท๎„‰๎„‰๎‚ญ๎„ซ๎ƒฐ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ฝ๎€„๎€œ๎๎€ง๎€ƒ๎๎๎๎ˆ˜๎€ƒ
While erm (42) gene alone was not found in any case, only msr (E)
resistance gene was found in 8 cases (4 calves, 2 young cattle, 2 adult
cattle) and only mph (E) gene was found in 2 cases (1 calf and 1 adult
cattle). While msr (E) and mph (E)๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ๎…˜๎„จ๎„ซ๎ƒ๎„ฎ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎ƒฆ๎ƒ๎„ด๎ƒฌ๎ƒ๎„ซ๎€ƒ๎ƒฐ๎„Š๎€ƒ
17 cases (7 calves, 7 young cattle, 3 adult cattle), three of msr (E), mph
(E) and erm (42)๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ๎…˜๎„จ๎„ซ๎ƒ๎„ฎ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎ƒฆ๎ƒ๎„ด๎ƒฌ๎ƒ๎„ซ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‡ ๎‡ก๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ˆง๎‡ค๎€ƒ๎ƒ†๎‚ญ๎„ƒ๎…’๎ƒ๎„ฎ๎ˆ™๎€ƒ
5 young cattle, 2 adult cattle). In 13 cases (4 calves, 8 young cattle, 1
adult cattle), no resistance gene was found (๎ฝ๎€„๎€œ๎๎€ง๎€ƒ๎๎๎๎ˆจ๎ˆ˜
๎ฝ๎ƒฌ๎ƒ๎„ซ๎ƒ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„ฎ๎„๎„‰๎ƒ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎ƒฐ๎ƒ๎„ฎ๎€ƒ๎„๎„Š๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„ฎ๎„ท๎„ฎ๎ƒ†๎ƒ๎„จ๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎…™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ
of M. haemolytica in our country and in the world. In the wide ranging
study of Portis et al. [8๎ˆฌ๎€ƒ๎„Š๎„๎€ƒ๎„ฎ๎ƒฐ๎ƒฆ๎„Š๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„Š๎„ด๎€ƒ๎ƒ†๎ƒฌ๎‚ญ๎„Š๎ƒฆ๎ƒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ
distribution of minimal inhibitory concentration (MIC) distributions
for penicillin and ceftiofur among M. haemolytica isolates, while
๎ƒŒ๎ƒ๎ƒ†๎„ซ๎ƒ๎‚ญ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎„ฎ๎„ท๎„ฎ๎ƒ†๎ƒ๎„จ๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎…™๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„๎ƒ…๎„ฎ๎ƒ๎„ซ๎…’๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎ƒ๎„Š๎„ซ๎„๎…ฆ๎„๎…˜๎‚ญ๎ƒ†๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ
group antibiotics tilmicosin and tulathromycin. As a result of this
study, it was emphasised that crossโ€“resistance occurred in macrolide
group antimicrobials. In presented study, M. haemolytica was detected
๎ƒฐ๎„Š๎€ƒ๎‡ค๎‡ก๎‰ฆ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ
observed in 75% of the cases.
In a study [10] conducted in healthy (n:107) and feedlot cattle with
pneumonic pasteurellosis (n:210), the prevalence and antimicrobial
susceptibility of three diseaseโ€“associated bacteria (M. haemolytica,
๎ฑ๎ˆ˜๎‰๎„‰๎„ท๎„ƒ๎„ด๎„๎ƒ†๎ƒฐ๎ƒŒ๎‚ญ and H. somni) were analyzed. In diseased cattle, P. multocida
was isolated most frequently (54.8%), followed by M. haemolytica (30.5%)
and H. somni (22.9%). It was reported that ๎”๎ˆ˜๎‰๎ƒฌ๎‚ญ๎ƒ๎„‰๎„๎„ƒ๎…™๎„ด๎ƒฐ๎ƒ†๎‚ญ was isolated
more frequently in healthy cattle compared to others. While high
resistance levels (>70%) to tulathromycin, one of the macrolide group
antibiotics, were determined in M. haemolytica isolates. Researchers
[10] stated that this resistance may be caused by macrolide resistance
genes [msr (E), mph (E), erm (42)], but additional studies should be
carried out in this regard.
All of the antibiotic susceptibility or resistance tests used in
these studies are based on the technique of measuring MIC values
of drugs by tube dilution or disc diffusion (Kirby Bauer) techniques
[17]. However, these tests have disadvantages such as the inability
to grow bacteria in culture medium from samples taken from living
๎„๎„ซ๎ƒฆ๎‚ญ๎„Š๎ƒฐ๎„ฎ๎„‰๎„ฎ๎€ƒ๎„ด๎„ซ๎ƒ๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎„ฎ๎€ƒ๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎‚ญ๎ƒ†๎„๎€ƒ๎„๎ƒฅ๎€ƒ๎„ฎ๎„ท๎…จ๎ƒ†๎ƒฐ๎ƒ๎„Š๎„ด๎€ƒ๎ƒ…๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎€ƒ
๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎„๎„Š๎ƒฆ๎€ƒ๎ƒŒ๎„ท๎„ซ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ†๎„ท๎„ƒ๎„ด๎„ท๎„ซ๎ƒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ฎ๎ƒ๎„Š๎„ฎ๎ƒฐ๎„ด๎ƒฐ๎…’๎ƒฐ๎„ด๎…™๎€ƒ๎„ด๎ƒ๎„ฎ๎„ด๎„ฎ๎ˆ˜๎€ƒ๎€น๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„ซ๎ƒ๎‚ญ๎„ฎ๎„๎„Š๎ˆ™๎€ƒ๎ƒฐ๎„ด๎€ƒ
is very important to determine antibiotic resistance genes in bacteria
by using PCR method, which is a shorter and more reliable test to
determine antibiotic resistance in bacteria.
In a study conducted by Michael et al. [18] to determine the macrolide
resistance genes in M. haemolytica, they determined erm (42) genes in 1
of 29 M. haemolytica isolates and erm (42)+msr (E)+mph (E) genes in 21 of
๎„ด๎ƒฌ๎ƒ๎„‰๎€ƒ๎ƒ…๎…™๎€ƒ๎ฑ๎€๎ด๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎…™๎€ƒ๎ƒฅ๎„๎„ท๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎”๎๎€๎€ƒ๎…’๎‚ญ๎„ƒ๎„ท๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎ƒ†๎‚ญ๎„ซ๎„ซ๎…™๎ƒฐ๎„Š๎ƒฆ๎€ƒ
these three genes increased by 32โ€“64 times for both gamithromycin
and tildiprosin, while the MIC value of gamithromycin increased by 4
times and the MIC value of tildiprosin increased by 32 times in those
with erm (42)๎€ƒ๎‚ญ๎„ƒ๎„๎„Š๎ƒ๎ˆ˜๎€ƒ๎ด๎„๎„ฎ๎ƒ๎‰๎ƒ๎„ด๎‰๎‚ญ๎„ƒ. [9] in their study to determine the
resistance to tulathromycin and tilmicosin, which are macrolides used
๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…’๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎‚ญ๎„ซ๎…™๎€ƒ๎…ฅ๎ƒ๎„ƒ๎ƒŒ๎ˆ™๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒฆ๎‚ญ๎„‰๎ƒฐ๎„ด๎ƒฌ๎„ซ๎„๎„‰๎…™๎ƒ†๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฐ๎„ƒ๎ƒŒ๎„๎„จ๎„ซ๎„๎„ฎ๎ƒฐ๎„Š๎ˆ™๎€ƒ๎…“๎ƒฌ๎ƒฐ๎ƒ†๎ƒฌ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ
new compounds; erm (42) in 6, msr (E) โ€“mph (E) in 5 and erm (42)+msr (E)
โ€“mph (E) in 4 of 20 M.haemolytica ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒ…๎…™๎€ƒ๎„‰๎„ท๎„ƒ๎„ด๎ƒฐ๎„จ๎„ƒ๎ƒ๎…˜๎€ƒ
PCR method. It was determined that the MIC values of tildiprosine,
๎„ด๎ƒฐ๎„ƒ๎„‰๎ƒฐ๎ƒ†๎„๎„ฎ๎ƒฐ๎„Š๎ˆ™๎€ƒ๎„ด๎„ท๎„ƒ๎‚ญ๎„ด๎ƒฌ๎„ซ๎„๎„‰๎…™๎ƒ†๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎ƒฆ๎‚ญ๎„‰๎ƒฐ๎„ด๎ƒฌ๎„ซ๎„๎„‰๎…™๎ƒ†๎ƒฐ๎„Š๎€ƒ๎ƒฐ๎„Š๎ƒ†๎„ซ๎ƒ๎‚ญ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎‡ง๎€ƒ๎ˆฏ๎€ƒ๎‰–๎‡ ๎‡ก๎‡ง๎€ƒ๎„ด๎ƒฐ๎„‰๎ƒ๎„ฎ๎€ƒ
in bacterial isolates containing only erm (42) or msr (E) โ€“mph (E) genes
๎„๎„ซ๎€ƒ๎‚ญ๎„ƒ๎„ƒ๎€ƒ๎„ด๎ƒฌ๎„ซ๎ƒ๎ƒ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒ๎„ด๎„๎ƒฆ๎ƒ๎„ด๎ƒฌ๎ƒ๎„ซ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎„ท๎„ฎ๎ˆ™๎€ƒ๎„ด๎ƒฌ๎ƒ๎…™๎€ƒ๎ƒŒ๎ƒ๎„‰๎„๎„Š๎„ฎ๎„ด๎„ซ๎‚ญ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎„ฎ๎ƒ๎€ƒ๎„ด๎ƒฌ๎„ซ๎ƒ๎ƒ๎€ƒ
genes [erm (42), msr (E), mph (E)] were responsible for resistance to
macrolide antibiotics in M.haemolytica isolates.
_____________________________________________________________________________Revista Cientifica, FCV-LUZ / Vol. XXXIV, rcfcv-e34423
7 of 8
As stated in all these studies, it is observed that erm (42), msr (E),
mph (E) genes increase the MIC values against macrolide antibiotics
and therefore they are responsible for the resistance against them. In
the present study, at least one of these resistance genes was detected
๎‚ญ๎„ฎ๎€ƒ๎‚ญ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„ท๎„ƒ๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎„‰๎„ท๎„ƒ๎„ด๎ƒฐ๎„จ๎„ƒ๎ƒ๎…˜๎€ƒ๎ฑ๎€๎ด๎€ƒ๎‚ญ๎„Š๎‚ญ๎„ƒ๎…™๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‡ฆ๎‡ค๎‰ฆ๎€ƒ๎„๎ƒฅ๎€ƒ๎‡ค๎‡ก๎€ƒ๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎ƒฅ๎„๎„ท๎„Š๎ƒŒ๎€ƒ๎„จ๎„๎„ฎ๎ƒฐ๎„ด๎ƒฐ๎…’๎ƒ๎€ƒ
for M. haemolytica๎€ƒ๎ƒ…๎…™๎€ƒ๎๎€พ๎€๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„ท๎„ƒ๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‚ญ๎„Š๎€ƒ๎ƒฐ๎„Š๎ƒ†๎„ซ๎ƒ๎‚ญ๎„ฎ๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ
the MIC values of macrolide group antibiotics to various degrees
during the treatment process and consequently, deaths occurred as
a result of the failure to achieve the desired success in treatment. It
is important to consider antimicrobial resistance and the spread of
antimicrobial resistance genes in treatment decisions associated with
common diseases such as BRD. Understanding the epidemiology of
antimicrobial resistance in BRD pathogens in healthy cattle, as well
as treated and untreated sick cattle, is crucial to understanding the
role of resistance in BRD treatment failure [19].
In these studies, it is also stated that the macrolide resistance
mechanism formed in bacteria may be caused by at least 3 different
๎ƒ†๎„๎„‰๎ƒ…๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒerm (42)๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎‚ญ๎„Š๎„ด๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„ซ๎„ฎ๎„ด๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ
๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎ˆ˜๎€ƒ๎๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎ˆ™๎€ƒ๎”๎๎ท
B
(macrolide, lincosamide and
streptogramin B) phenotype with high resistance to lincosamides
and low to medium resistance to macrolide and streptogramin B
๎ƒฐ๎„ฎ๎€ƒ๎ƒฅ๎„๎„ซ๎„‰๎ƒ๎ƒŒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎ƒ๎ƒ†๎„๎„Š๎ƒŒ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎„ด๎€ƒ
without lincosamide resistance and contained two resistance genes,
๎„‰๎„ฎ๎„ซ๎‰๎ˆง๎€ง๎ˆจ and mph (E),๎€ƒ๎ƒ๎„Š๎ƒ†๎„๎ƒŒ๎ƒฐ๎„Š๎ƒฆ๎€ƒ๎‚ญ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎ƒ๎…ฉ๎„ท๎…˜๎€ƒ๎„จ๎„ท๎„‰๎„จ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ
๎ƒฐ๎„Š๎‚ญ๎ƒ†๎„ด๎ƒฐ๎…’๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„จ๎ƒฌ๎„๎„ฎ๎„จ๎ƒฌ๎„๎„ด๎„ซ๎‚ญ๎„Š๎„ฎ๎ƒฅ๎ƒ๎„ซ๎‚ญ๎„ฎ๎ƒ๎ˆ™๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎ƒ๎ƒ†๎„ด๎ƒฐ๎…’๎ƒ๎„ƒ๎…™๎ˆ™๎€ƒ๎ƒ๎…˜๎„จ๎„ซ๎ƒ๎„ฎ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ
๎„ฎ๎‚ญ๎„‰๎ƒ๎€ƒ๎„ฎ๎„๎„ท๎„ซ๎ƒ†๎ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ซ๎ƒŒ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎ƒฌ๎ƒฐ๎ƒฆ๎ƒฌ๎„ƒ๎…™๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎„ด๎€ƒ๎„ด๎„๎€ƒ๎‚ญ๎€ƒ
comprehensive set of macrolides and includes all three determinants
erm (42), msr (E) and mph (E) [7, 8]. Regarding these issues, Owen et al.
[19] stated that their study showed that antimicrobial susceptibility
phenotypes are necessary to complement genomically predicted
antibiotic resistance gene genotypes in order to better understand
how the presence of antibiotic resistance genes in a particular
bacterial species can potentially affect optimal treatment of bovine
respiratory disease and morbidity/mortality outcomes.
๎€น๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ๎„จ๎„๎ƒฐ๎„Š๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎…’๎ƒฐ๎ƒ๎…“๎ˆ™๎€ƒ๎„Š๎„๎€ƒ๎ƒฐ๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒ๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎ƒฅ๎„๎„ท๎„Š๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„ซ๎„ฎ๎„ด๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎ƒฐ๎„Š๎€ƒ
which only erm (42)๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎ƒฅ๎„๎„ท๎„Š๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„จ๎„ซ๎ƒ๎„ฎ๎ƒ๎„Š๎„ด๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ฎ๎ƒ๎ƒ†๎„๎„Š๎ƒŒ๎€ƒ๎ƒฆ๎„ซ๎„๎„ท๎„จ๎ˆ™๎€ƒ
i.e. 17 isolates with msr (E) and mph (E) genes together (7 calves, 7
young cattle, 3 adult cattle), and the third group, in which msr (E),
mph (E) and erm (42) genes were found together, was 12 (5 calves, 5
๎…™๎„๎„ท๎„Š๎ƒฆ๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎ˆ™๎€ƒ๎‡ก๎€ƒ๎‚ญ๎ƒŒ๎„ท๎„ƒ๎„ด๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎ˆจ๎€ƒ๎ˆง๎ฝ๎€„๎€œ๎๎€ง๎€ƒ๎๎๎๎ˆจ๎ˆ˜๎€ƒ๎๎„ด๎€ƒ๎…“๎‚ญ๎„ฎ๎€ƒ๎„ซ๎ƒ๎„จ๎„๎„ซ๎„ด๎ƒ๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎”๎๎€๎€ƒ
value of tulathromycin, gamithromycin, tilmicosin and clindamycin
increased 16โ€“128 times in group 2 isolates and 128 โ€“ >1024 times in
group 3 isolates [7]. According to Alhamami et al. [20] noting that
most of the isolates were obtained from BRDโ€“affected animals with
a history of antimicrobial therapy, they suggested possible treatment
๎ƒฅ๎‚ญ๎ƒฐ๎„ƒ๎„ท๎„ซ๎ƒ๎€ƒ๎ƒŒ๎„ท๎ƒ๎€ƒ๎„ด๎„๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„ท๎„ฎ๎‚ญ๎„ด๎ƒฐ๎…’๎ƒ๎€ƒ๎„จ๎‚ญ๎„ด๎ƒฌ๎„๎ƒฆ๎ƒ๎„Š๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒ๎‚ญ๎„ซ๎ƒ†๎ƒฌ๎ƒ๎„ซ๎„ฎ๎€ƒ
also suggested that continuous monitoring is necessary to support
the antimicrobial management programs recently developed by
the industry to ensure the effectiveness of veterinary antimicrobial
๎ƒŒ๎„ซ๎„ท๎ƒฆ๎„ฎ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎„ฎ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„ท๎„ƒ๎„ด๎„ฎ๎€ƒ๎„ฎ๎ƒฌ๎„๎…“๎€ƒ๎ƒฌ๎„๎…“๎€ƒ๎ƒฐ๎„‰๎„จ๎„๎„ซ๎„ด๎‚ญ๎„Š๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„จ๎„ซ๎ƒ๎„ฎ๎ƒ๎„Š๎ƒ†๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ
genes is in treatment.
CONCLUSION
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎ƒ๎„ด๎ƒฐ๎„๎„ƒ๎„๎ƒฆ๎ƒฐ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒM. haemolytica by IHC method in
only fibrinous pneumonia cases in Konya and its surrounding
provinces and the presence of resistance genes against macrolide
๎ƒฆ๎„ซ๎„๎„ท๎„จ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎„ฎ๎€ƒ๎…“๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎ƒ๎ƒŒ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎„ซ๎„ฎ๎„ด๎€ƒ๎„ด๎ƒฐ๎„‰๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆ˜๎€ƒ๎๎„ด๎€ƒ๎ƒฐ๎„ฎ๎€ƒ
thought that the use of IHC method in cases where fresh tissue is
not available, autolysis and antimicrobial treatment is applied makes
๎‚ญ๎€ƒ๎„ฎ๎ƒฐ๎ƒฆ๎„Š๎ƒฐ๎…ฅ๎ƒ†๎‚ญ๎„Š๎„ด๎€ƒ๎ƒ†๎„๎„Š๎„ด๎„ซ๎ƒฐ๎ƒ…๎„ท๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒ๎„ด๎ƒฐ๎„๎„ƒ๎„๎ƒฆ๎ƒฐ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒฌ๎„ท๎„ฎ๎€ƒ๎„ด๎ƒฌ๎ƒฐ๎„ฎ๎€ƒ
method can be safely transferred to routine studies, especially in
๎ƒ†๎‚ญ๎„ฎ๎ƒ๎„ฎ๎€ƒ๎…“๎ƒฌ๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒ…๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒฐ๎„๎„ƒ๎„๎ƒฆ๎ƒฐ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎‚ญ๎„Š๎‚ญ๎„ƒ๎…™๎„ฎ๎ƒ๎„ฎ๎€ƒ๎‚ญ๎„ซ๎ƒ๎€ƒ๎„Š๎„๎„ด๎€ƒ๎‚ญ๎„จ๎„จ๎„ซ๎„๎„จ๎„ซ๎ƒฐ๎‚ญ๎„ด๎ƒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎„ซ๎ƒ๎€ƒ๎„‰๎‚ญ๎…™๎€ƒ
๎ƒ…๎ƒ๎€ƒ๎„‰๎‚ญ๎„Š๎…™๎€ƒ๎„ซ๎ƒ๎‚ญ๎„ฎ๎„๎„Š๎„ฎ๎€ƒ๎ƒฅ๎„๎„ซ๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒฅ๎‚ญ๎ƒฐ๎„ƒ๎„ท๎„ซ๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎„ด๎„ซ๎ƒ๎‚ญ๎„ด๎„‰๎ƒ๎„Š๎„ด๎ˆ˜๎€ƒ
It was concluded that one of these reasons may be M. haemolytica
isolates containing erm (42), msr (E), mph (E) genes responsible for
resistance to macrolide antibiotics commonly used in the treatment
๎„๎ƒฅ๎€ƒ๎…ฅ๎ƒ…๎„ซ๎ƒฐ๎„Š๎„๎„ท๎„ฎ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎ˆ˜๎€ƒ๎๎„ด๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎„๎„Š๎„๎…“๎„Š๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎„ด๎ƒฌ๎ƒ๎„ฎ๎ƒ๎€ƒ๎ƒฆ๎ƒ๎„Š๎ƒ๎„ฎ๎€ƒ๎ƒ†๎‚ญ๎„Š๎€ƒ๎ƒ…๎ƒ๎€ƒ๎ƒฅ๎„๎„ท๎„Š๎ƒŒ๎€ƒ
regardless of age or disease/health status, that they are responsible
for crossโ€“resistance between this group of antibiotics, and that
resistance may even develop against macrolide group antibiotics that
๎ƒฌ๎‚ญ๎…’๎ƒ๎€ƒ๎„Š๎„๎„ด๎€ƒ๎…™๎ƒ๎„ด๎€ƒ๎ƒ…๎ƒ๎ƒ๎„Š๎€ƒ๎ƒฐ๎„Š๎„ด๎„ซ๎„๎ƒŒ๎„ท๎ƒ†๎ƒ๎ƒŒ๎€ƒ๎„ด๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎…ฅ๎ƒ๎„ƒ๎ƒŒ๎ˆ˜๎€ƒ๎ฝ๎ƒฌ๎ƒ๎„ซ๎ƒ๎ƒฅ๎„๎„ซ๎ƒ๎ˆ™๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ƒฅ๎„ซ๎ƒ๎„ช๎„ท๎ƒ๎„Š๎ƒ†๎…™๎€ƒ
๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„ฎ๎„ท๎„ฎ๎ƒ†๎ƒ๎„จ๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎…™๎€ƒ๎„จ๎„ซ๎„๎…ฅ๎„ƒ๎ƒ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒM. haemolytica should be
periodically renewed due to the possible development of resistance
as it will help to guide the choice of antimicrobials in the control and
treatment of such diseases. As can be seen from the presented study,
repeated and unlimited use of these antibiotics should be avoided in
cases where the frequency of resistance is high.
ACKNOWLEDGMENT
๎ฝ๎ƒฌ๎ƒ๎€ƒ๎‚ญ๎„ท๎„ด๎ƒฌ๎„๎„ซ๎„ฎ๎€ƒ๎ƒฆ๎„ซ๎‚ญ๎„ด๎ƒ๎ƒฅ๎„ท๎„ƒ๎„ƒ๎…™๎€ƒ๎„ด๎ƒฌ๎‚ญ๎„Š๎„๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎ท๎ƒ๎„ƒ๎ƒ†๎„ท๎„๎€ƒ๎‚€๎„Š๎ƒฐ๎…’๎ƒ๎„ซ๎„ฎ๎ƒฐ๎„ด๎…™๎€ƒ๎ท๎ƒ†๎ƒฐ๎ƒ๎„Š๎„ด๎ƒฐ๎ƒฅ๎ƒฐ๎ƒ†๎€ƒ
๎ด๎ƒ๎„ฎ๎ƒ๎‚ญ๎„ซ๎ƒ†๎ƒฌ๎€ƒ๎ฑ๎„ซ๎„๎ƒพ๎ƒ๎ƒ†๎„ด๎„ฎ๎€ƒ๎ˆง๎€œ๎€„๎ฑ๎ˆจ๎€ƒ๎€๎„๎„๎„ซ๎ƒŒ๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎š๎…จ๎ƒ†๎ƒ๎€ƒ๎ˆง๎ฑ๎„ซ๎„๎ƒพ๎ƒ๎ƒ†๎„ด๎€ƒ๎•๎„๎ˆš๎€ƒ๎‡ ๎‡ฆ๎‡ฃ๎‡Ÿ๎‡ ๎‡Ÿ๎‡ค๎‡ฅ๎ˆจ๎€ƒ
๎ƒฅ๎„๎„ซ๎€ƒ๎ƒฅ๎„ท๎„Š๎ƒŒ๎ƒถ๎„Š๎ƒฆ๎€ƒ๎„ด๎ƒฌ๎ƒถ๎„ฎ๎€ƒ๎„ฎ๎„ด๎„ท๎ƒŒ๎…™๎ˆ˜
Con๎…ฆict of interest
๎ฝ๎ƒฌ๎ƒ๎„ซ๎ƒ๎€ƒ๎ƒฐ๎„ฎ๎€ƒ๎„Š๎„๎€ƒ๎ƒ†๎„๎„Š๎…ฆ๎ƒฐ๎ƒ†๎„ด๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฐ๎„Š๎„ด๎ƒ๎„ซ๎ƒ๎„ฎ๎„ด๎€ƒ๎ƒ…๎ƒ๎„ด๎…“๎ƒ๎ƒ๎„Š๎€ƒthe authors.
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๎ˆซ๎‡ข๎ˆฌ๎€ƒ ๎•๎ƒฆ๎„ท๎…™๎ƒ๎„Š๎€ƒ๎ฑ๎‚“๎ˆ™๎€ƒ๎๎ƒ๎€ƒ๎€๎ฝ๎ˆ™๎€ƒ๎€พ๎„๎€ƒ๎‚™๎ฝ๎ฝ๎ˆ™๎€ƒ๎ฝ๎„ซ๎„ท๎„๎„Š๎ƒฆ๎€ƒ๎ฑ๎€พ๎ˆ™๎€ƒ๎๎„๎ƒฐ๎€ƒ๎€œ๎‚“๎ˆ™๎€ƒ๎•๎ƒฆ๎„ท๎…™๎ƒ๎„Š๎€ƒ๎Ž๎€๎ฝ๎ˆ˜๎€ƒ๎€น๎ƒฐ๎„ซ๎„ฎ๎„ด๎€ƒ
report of antimicrobial resistance of Mannheimia haemolytica from
๎ฑ๎ƒฌ๎‚ญ๎„Š๎€ƒ๎ด๎‚ญ๎„Š๎ƒฆ๎€ƒ๎„ฎ๎ƒฌ๎ƒ๎ƒ๎„จ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎‚“๎ƒฐ๎ƒ๎„ด๎„Š๎‚ญ๎„‰๎ˆ˜๎€ƒ๎ฑ๎‚ญ๎„๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ๎Œ๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ข๎€ƒ๎ˆซ๎ƒ†๎ƒฐ๎„ด๎ƒ๎ƒŒ๎€ƒ
๎‡ก๎‡ง๎€ƒ๎Œ๎„ท๎„ƒ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ฃ๎ˆฌ๎ˆ›๎€ƒ๎‡ฃ๎‡ข๎ˆง๎‡ ๎ˆจ๎ˆš๎‡ฃ๎‡ ๎ˆฏ๎‡ฃ๎‡ง๎ˆ˜๎€ƒ๎€„๎…’๎‚ญ๎ƒฐ๎„ƒ๎‚ญ๎ƒ…๎„ƒ๎ƒ๎€ƒ๎ƒฐ๎„Š๎ˆš๎€ƒ๎ƒฌ๎„ด๎„ด๎„จ๎„ฎ๎ˆš๎ˆฅ๎ˆฅ๎ƒฆ๎„๎„๎ˆ˜๎„ฎ๎„ท๎ˆฅ๎„ซ๎…ก๎‡ฆ๎ƒ…๎€๎€พ๎ฝ
๎ˆซ๎‡ฃ๎ˆฌ๎€ƒ ๎๎„๎„จ๎ƒ๎…ก๎€ƒ๎€„๎ˆ™๎€ƒ๎”๎‚ญ๎„ซ๎„ด๎ƒฐ๎„Š๎„ฎ๎„๎„Š๎€ƒ๎ท๎€„๎ˆ˜๎€ƒ๎ด๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎„ฎ๎…™๎„ฎ๎„ด๎ƒ๎„‰๎ˆ™๎€ƒ๎„‰๎ƒ๎ƒŒ๎ƒฐ๎‚ญ๎„ฎ๎„ด๎ƒฐ๎„Š๎„ท๎„‰๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ
๎„จ๎„ƒ๎ƒ๎„ท๎„ซ๎‚ญ๎ƒ๎ˆ˜๎€ƒ๎๎„Š๎ˆš๎€ƒ๎‚ข๎‚ญ๎ƒ†๎ƒฌ๎‚ญ๎„ซ๎…™๎€ƒ๎Œ๎€น๎ˆ™๎€ƒ๎ƒ๎ƒŒ๎ƒฐ๎„ด๎„๎„ซ๎ˆ˜๎€ƒ๎ฑ๎‚ญ๎„ด๎ƒฌ๎„๎„ƒ๎„๎ƒฆ๎ƒฐ๎ƒ†๎€ƒ๎ƒ…๎‚ญ๎„ฎ๎ƒฐ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎…’๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎‚ญ๎„ซ๎…™๎€ƒ
disease. 6
th
ed. Maryland heights (Missouri, USA): Mosby; 2017.
p.471โ€“560.
๎ˆซ๎‡ค๎ˆฌ๎€ƒ ๎‚€๎ƒŒ๎ƒŒ๎ƒฐ๎„Š๎€ƒ๎ฝ๎”๎ˆ™๎€ƒ๎€๎ƒฌ๎‚ญ๎„๎„ซ๎‚ญ๎ƒ…๎„๎„ซ๎„ด๎…™๎€ƒ๎€„๎Œ๎ˆ™๎€ƒ๎Ž๎ƒฌ๎„ท๎„ฎ๎„ซ๎„๎€ƒ๎€„๎ˆ™๎€ƒ๎‚ข๎ƒฐ๎ƒŒ๎‚ญ๎„Š๎€ƒ๎€œ๎ด๎”๎ˆ™๎€ƒ๎”๎ƒฐ๎„ด๎„ซ๎‚ญ๎€ƒ๎ท๎ˆ™๎€ƒ๎€ง๎„‰๎„ซ๎‚ญ๎„Š๎€ƒ
๎ฝ๎€œ๎ˆ™๎€ƒ๎€ฃ๎ƒฌ๎‚ญ๎„‰๎‚ญ๎€ƒ๎Ž๎ˆ™๎€ƒ๎ด๎ƒฐ๎„จ๎„๎„Š๎€ƒ๎”๎Ž๎€พ๎ˆ™๎€ƒ๎€บ๎‚ญ๎ƒพ๎ƒŒ๎‚ฎ๎ƒ†๎„ฎ๎€ƒ๎”๎ˆ™๎€ƒ๎ท๎‚ญ๎ƒฌ๎ƒฐ๎ƒ…๎…ก๎‚ญ๎ƒŒ๎‚ญ๎€ƒ๎”๎‚€๎Ž๎ˆ™๎€ƒ๎€พ๎„๎„ฎ๎„ฎ๎‚ญ๎ƒฐ๎„Š๎€ƒ
๎”๎Œ๎ˆ™๎€ƒ๎Ž๎„๎ƒฐ๎„ซ๎‚ญ๎„ƒ๎‚ญ๎€ƒ๎•๎ˆ˜๎€ƒ๎€„๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ƒ๎„ฎ๎ˆš๎€ƒ๎€พ๎ƒฐ๎„ฎ๎„ด๎„๎„ซ๎…™๎ˆ™๎€ƒ
mechanisms, therapeutic strategies and future prospects.
๎Œ๎ˆ˜๎€ƒ๎๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ˆ˜๎€ƒ๎ฑ๎„ท๎ƒ…๎„ƒ๎ƒฐ๎ƒ†๎€ƒ๎€พ๎ƒ๎‚ญ๎„ƒ๎„ด๎ƒฌ๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ ๎ˆ›๎€ƒ๎‡ ๎‡ฃ๎ˆง๎‡ ๎‡ก๎ˆจ๎ˆš๎‡ ๎‡ฆ๎‡ค๎‡Ÿ๎ˆฏ๎‡ ๎‡ฆ๎‡ฅ๎‡ฅ๎ˆ˜๎€ƒ๎ƒŒ๎„๎ƒฐ๎ˆš๎€ƒ
https://doi.org/gsdt63
๎ˆซ๎‡ฅ๎ˆฌ๎€ƒ ๎€พ๎‚ญ๎„‰๎„‰๎„๎„Š๎ƒŒ๎€ƒ๎ด๎Œ๎€พ๎ˆ™๎€ƒ๎€น๎‚ญ๎„ƒ๎ƒ†๎„๎„Š๎ƒ๎„ซ๎€ƒ๎Ž๎ˆ™๎€ƒ๎ฑ๎„๎…“๎ƒ๎„ƒ๎„ƒ๎€ƒ๎ฝ๎ˆ™๎€ƒ๎€œ๎„๎…“๎„Š๎ƒ๎„ฎ๎„ฎ๎€ƒ๎ด๎ˆ™๎€ƒ๎€บ๎ƒฐ๎„ƒ๎„ƒ๎ƒ๎„ฎ๎„จ๎ƒฐ๎ƒ๎€ƒ๎ท๎€พ๎ˆ˜๎€ƒ
A simple labelโ€“free method reveals bacterial growth dynamics
and antibiotic action in realโ€“time. Sci. Rep. [Internet]. 2022;
12(19393). doi: https://doi.org/g8wtk7
Mannheimia haemolytica resistance genes of cattle / Hatipoglu et al. ______________________________________________________________
8 of 8
[7] Desmolaize B, Rose S, Wilhelm C, Warrass R, Douthwaite S.
๎€๎„๎„‰๎ƒ…๎ƒฐ๎„Š๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎‚ญ๎„Š๎„ด๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎…ฅ๎ƒ๎„ƒ๎ƒŒ๎€ƒ
isolates of Mannheimia haemolytica and Pasteurella multocida.
Antimicrob. Agents Chemother. [Internet]. 2011; 55(9):4128โ€“4133.
doi: https://doi.org/cgdh5p
๎ˆซ๎‡ง๎ˆฌ๎€ƒ ๎ฑ๎„๎„ซ๎„ด๎ƒฐ๎„ฎ๎€ƒ๎€ง๎ˆ™๎€ƒ๎๎ƒฐ๎„Š๎ƒŒ๎ƒ๎„‰๎‚ญ๎„Š๎€ƒ๎€๎ˆ™๎€ƒ๎Œ๎„๎ƒฌ๎‚ญ๎„Š๎„ฎ๎ƒ๎„Š๎€ƒ๎๎ˆ™๎€ƒ๎ท๎„ด๎„๎„ƒ๎„ด๎„‰๎‚ญ๎„Š๎€ƒ๎€บ๎ˆ˜๎€ƒ๎€„๎€ƒ๎„ด๎ƒ๎„Š๎ˆฏ๎…™๎ƒ๎‚ญ๎„ซ๎€ƒ
(2000โ€“2009) study of antimicrobial susceptibility of bacteria
๎„ด๎ƒฌ๎‚ญ๎„ด๎€ƒ๎ƒ†๎‚ญ๎„ท๎„ฎ๎ƒ๎€ƒ๎ƒ…๎„๎…’๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎ƒŒ๎ƒฐ๎„ฎ๎ƒ๎‚ญ๎„ฎ๎ƒ๎€ƒ๎ƒ†๎„๎„‰๎„จ๎„ƒ๎ƒ๎…˜๎ˆฏMannheimia
haemolytica, Pasteurella multocida, and Histophilus somniโ€“in
๎„ด๎ƒฌ๎ƒ๎€ƒ๎‚€๎„Š๎ƒฐ๎„ด๎ƒ๎ƒŒ๎€ƒ๎ท๎„ด๎‚ญ๎„ด๎ƒ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎€๎‚ญ๎„Š๎‚ญ๎ƒŒ๎‚ญ๎ˆ˜๎€ƒ๎Œ๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ๎€ฃ๎ƒฐ๎‚ญ๎ƒฆ๎ˆ˜๎€ƒ๎๎„Š๎…’๎ƒ๎„ฎ๎„ด๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ
2012; 24(5):932โ€“944. doi: https://doi.org/f39fr4
๎ˆซ๎‡จ๎ˆฌ๎€ƒ ๎ด๎„๎„ฎ๎ƒ๎€ƒ๎ท๎ˆ™๎€ƒ๎€ฃ๎ƒ๎„ฎ๎„‰๎„๎„ƒ๎‚ญ๎ƒฐ๎…ก๎ƒ๎€ƒ๎€œ๎ˆ™๎€ƒ๎Œ๎‚ญ๎ƒพ๎„ท๎€ƒ๎ฑ๎ˆ™๎€ƒ๎‚”๎ƒฐ๎„ƒ๎ƒฌ๎ƒ๎„ƒ๎„‰๎€ƒ๎€๎ˆ™๎€ƒ๎‚”๎‚ญ๎„ซ๎„ซ๎‚ญ๎„ฎ๎„ฎ๎€ƒ๎ด๎ˆ™๎€ƒ๎€ฃ๎„๎„ท๎„ด๎ƒฌ๎…“๎‚ญ๎ƒฐ๎„ด๎ƒ๎€ƒ
๎ท๎ˆ˜๎€ƒ๎”๎„ท๎„ƒ๎„ด๎ƒฐ๎„จ๎„ƒ๎ƒ๎…˜๎€ƒ๎ฑ๎€๎ด๎€ƒ๎„ด๎„๎€ƒ๎ƒฐ๎ƒŒ๎ƒ๎„Š๎„ด๎ƒฐ๎ƒฅ๎…™๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒŒ๎ƒ๎„ด๎ƒ๎„ซ๎„‰๎ƒฐ๎„Š๎‚ญ๎„Š๎„ด๎„ฎ๎€ƒ๎ƒฐ๎„Š๎€ƒ
Mannheimia haemolytica and Pasteurella multocida. Antimicrob.
Agents Chemother. [Internet]. 2012; 56(7):3664โ€“3669. doi:
https://doi.org/f32t33
๎ˆซ๎‡ ๎‡Ÿ๎ˆฌ๎€ƒ ๎ฝ๎ƒฐ๎„‰๎„ฎ๎ƒฐ๎„ด๎€ƒ๎€ง๎ˆ™๎€ƒ๎€พ๎‚ญ๎„ƒ๎„ƒ๎ƒ๎…“๎ƒ๎„ƒ๎„ƒ๎€ƒ๎Œ๎ˆ™๎€ƒ๎€œ๎„๎„๎„๎ƒ๎„ซ๎€ƒ๎€๎ˆ™๎€ƒ๎ฝ๎ƒฐ๎„ฎ๎„๎„Š๎€ƒ๎•๎ˆ™๎€ƒ๎€„๎„‰๎‚ญ๎„ด๎€ƒ๎ท๎ˆ™๎€ƒ๎€„๎„ƒ๎ƒ๎…˜๎‚ญ๎„Š๎ƒŒ๎ƒ๎„ซ๎€ƒ
๎ฝ๎‚”๎ˆ˜๎€ƒ๎ฑ๎„ซ๎ƒ๎…’๎‚ญ๎„ƒ๎ƒ๎„Š๎ƒ†๎ƒ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎„ฎ๎„ท๎„ฎ๎ƒ†๎ƒ๎„จ๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎…™๎€ƒ๎„๎ƒฅ๎€ƒMannheimia
haemolytica, Pasteurella multocida, and Histophilus somni isolated
from the lower respiratory tract of healthy feedlot cattle and
๎„ด๎ƒฌ๎„๎„ฎ๎ƒ๎€ƒ๎ƒŒ๎ƒฐ๎‚ญ๎ƒฆ๎„Š๎„๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎ƒ…๎„๎…’๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎ƒŒ๎ƒฐ๎„ฎ๎ƒ๎‚ญ๎„ฎ๎ƒ๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ๎”๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ƒฐ๎„๎„ƒ๎ˆ˜๎€ƒ
[Internet]. 2017; 208:118โ€“125. doi: https://doi.org/g8wtk8
๎ˆซ๎‡ ๎‡ ๎ˆฌ๎€ƒ ๎€ ๎ƒฐ๎ƒฅ๎„ด๎ƒ‰๎ƒฐ๎€ƒ๎”๎Ž๎ˆ™๎€ƒ๎š๎„ซ๎„ด๎‚ญ๎„ด๎‚ญ๎„ด๎„ƒ๎ƒฐ๎€ƒ๎”๎ˆ™๎€ƒ๎€ง๎„ซ๎ƒ๎„ซ๎€ƒ๎€พ๎ˆ™๎€ƒ๎€พ๎‚ญ๎„ด๎ƒฐ๎„จ๎„๎ƒง๎„ƒ๎„ท๎€ƒ๎€น๎ˆ™๎€ƒ๎š๎…ก๎ƒŒ๎ƒ๎„‰๎ƒฐ๎„ซ๎€ƒ๎š๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎ƒ๎„ซ๎€ƒ
๎ท๎ƒฐ๎„ฎ๎„ด๎ƒ๎„‰๎ƒฐ๎„๎€ƒ๎ฑ๎‚ญ๎„ด๎„๎„ƒ๎„๎ƒพ๎ƒฐ๎€ƒ๎๎€ƒ๎€๎ƒฐ๎„ƒ๎„ด๎€ƒ๎ˆง๎ท๎ƒฐ๎„Š๎ƒŒ๎ƒฐ๎„ซ๎ƒฐ๎„‰๎ˆฏ๎ท๎„๎„ƒ๎„ท๎„Š๎„ท๎„‰๎ˆจ๎€ƒ๎ˆซ๎‚“๎ƒ๎„ด๎ƒ๎„ซ๎ƒฐ๎„Š๎‚ญ๎„ซ๎…™๎€ƒ๎ท๎…™๎„ฎ๎„ด๎ƒ๎„‰๎ƒฐ๎ƒ†๎€ƒ
๎ฑ๎‚ญ๎„ด๎ƒฌ๎„๎„ƒ๎„๎ƒฆ๎…™๎ˆ˜๎€ƒ๎‚“๎„๎„ƒ๎ˆ˜๎€ƒ๎‡ ๎ˆ™๎€ƒ๎€ฃ๎ƒฐ๎ƒฆ๎ƒ๎„ฎ๎„ด๎ƒฐ๎…’๎ƒ๎ˆฏ๎ด๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎ˆจ๎ˆฌ๎ˆ˜๎€ƒ๎‡ฅ
th
ed. Istanbul
๎ˆง๎ฝ๎„ผ๎„ซ๎„๎ƒฐ๎…™๎ƒ๎ˆจ๎ˆš๎€ƒ๎•๎„๎ƒ…๎ƒ๎„ƒ๎€ƒ๎€„๎„๎‚ญ๎ƒŒ๎ƒ๎„‰๎ƒฐ๎„๎€ƒ๎‚š๎‚ญ๎…™๎ƒฐ๎„Š๎ƒ†๎ƒฐ๎„ƒ๎ƒฐ๎„๎ˆ›๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ข๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ฅ๎€ƒ๎„จ๎ˆ˜๎€ƒ๎ฝ๎„ท๎„ซ๎„๎ƒฐ๎„ฎ๎ƒฌ๎ˆ˜
๎ˆซ๎‡ ๎‡ก๎ˆฌ๎€ƒ ๎€พ๎‚ญ๎…ก๎ƒฐ๎„ซ๎„๎ƒฆ๎„ƒ๎„ท๎€ƒ๎ด๎ˆ™๎€ƒ๎€ง๎„ซ๎ƒŒ๎ƒ๎ƒฆ๎ƒ๎„ซ๎€ƒ๎Œ๎ˆ™๎€ƒ๎€บ๎„ท๎„ƒ๎ƒ…๎‚ญ๎ƒฌ๎‚ญ๎„ซ๎€ƒ๎”๎ˆ™๎€ƒ๎Ž๎„ท๎„ƒ๎€ƒ๎š๎ˆ˜๎€ƒ๎€„๎„ฎ๎„ฎ๎„๎ƒ†๎ƒฐ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ
Pasteurella haemolytica, Pasteurella multocida and Haemophilus
somnus๎€ƒ๎…“๎ƒฐ๎„ด๎ƒฌ๎€ƒ๎„จ๎„Š๎ƒ๎„ท๎„‰๎„๎„Š๎ƒฐ๎‚ญ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎ƒ†๎‚ญ๎„ƒ๎…’๎ƒ๎„ฎ๎ˆ˜๎€ƒ๎€ฃ๎„ด๎„ฎ๎ƒ†๎ƒฌ๎ˆ˜๎€ƒ๎ฝ๎ƒฐ๎ƒ๎„ซ๎‚ญ๎„ซ๎…ก๎„ด๎„ƒ๎ˆ˜๎€ƒ๎‚”๎„๎ƒ†๎ƒฌ๎ƒ๎„Š๎„ฎ๎ƒ†๎ƒฌ๎„ซ๎ˆ˜๎€ƒ
1997;104(4):150โ€“153. PMID: 9151475.
๎ˆซ๎‡ ๎‡ข๎ˆฌ๎€ƒ ๎Ž๎ƒฐ๎„ƒ๎ƒฐ๎ƒ†๎€ƒ๎€„๎ˆ™๎€ƒ๎”๎„ท๎…ก๎€ƒ๎€„๎ˆ˜๎€ƒ๎ฑ๎„Š๎„˜๎„‰๎„๎„Š๎ƒฐ๎„ƒ๎ƒฐ๎€ƒ๎„ฎ๎ƒฑ๎ƒง๎ƒฑ๎„ซ๎€ƒ๎‚ญ๎„๎ƒ†๎ƒฐ๎ƒง๎ƒ๎„ซ๎„ƒ๎ƒ๎„ซ๎ƒฐ๎„Š๎ƒŒ๎ƒ๎„Š๎€ƒ๎ƒ…๎‚ญ๎„๎„ด๎ƒ๎„ซ๎ƒฐ๎€ƒ๎ƒฐ๎…ก๎‚ญ๎„ƒ๎„๎„ฎ๎…™๎‚ญ๎„Š๎„ƒ๎‚ญ๎„ซ๎ƒฑ๎€ƒ
ve izole Pasteurella๎ˆธ๎„ƒ๎‚ญ๎„ซ๎ƒฑ๎„Š๎€ƒ๎ฑ๎„๎„ƒ๎ƒฐ๎„‰๎ƒ๎„ซ๎‚ญ๎…ก๎€ƒ๎‚ข๎ƒฐ๎„Š๎ƒ†๎ƒฐ๎„ซ๎€ƒ๎ด๎ƒ๎‚ญ๎„๎„ฎ๎ƒฐ๎…™๎„๎„Š๎„ท๎€ƒ๎ˆง๎ฑ๎‚ข๎ด๎ˆจ๎€ƒ๎ƒฐ๎„ƒ๎ƒ๎€ƒ
๎„ฎ๎‚ญ๎„จ๎„ด๎‚ญ๎„Š๎„‰๎‚ญ๎„ฎ๎ƒฑ๎€ƒ๎ˆซ๎๎„ฎ๎„๎„ƒ๎‚ญ๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ…๎‚ญ๎ƒ†๎„ด๎ƒ๎„ซ๎ƒฐ๎‚ญ๎„ƒ๎€ƒ๎‚ญ๎ƒฆ๎ƒ๎„Š๎„ด๎„ฎ๎€ƒ๎ƒฅ๎„ซ๎„๎„‰๎€ƒ๎„ด๎ƒฌ๎ƒ๎€ƒ๎„ƒ๎„ท๎„Š๎ƒฆ๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎€ƒ
with pneumonia and detection of Pasteurella spp. by Polymerase
๎€๎ƒฌ๎‚ญ๎ƒฐ๎„Š๎€ƒ๎ด๎ƒ๎‚ญ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎ˆฌ๎ˆ˜๎€ƒ๎ฝ๎„ท๎„ซ๎„๎ˆ˜๎€ƒ๎Œ๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ๎€„๎„Š๎ƒฐ๎„‰๎ˆ˜๎€ƒ๎ท๎ƒ†๎ƒฐ๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡Ÿ๎‡ฃ๎€ƒ๎ˆซ๎ƒ†๎ƒฐ๎„ด๎ƒ๎ƒŒ๎€ƒ๎‡ ๎‡ก๎€ƒ
๎Œ๎„ท๎„ƒ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ฃ๎ˆฌ๎ˆ›๎€ƒ๎‡ก๎‡ง๎ˆง๎‡ ๎ˆจ๎ˆš๎‡ก๎‡ ๎‡ฆ๎ˆฏ๎‡ก๎‡ก๎‡ข๎ˆ˜๎€ƒ๎ฝ๎„ท๎„ซ๎„๎ƒฐ๎„ฎ๎ƒฌ๎ˆ˜๎€ƒ๎€„๎…’๎‚ญ๎ƒฐ๎„ƒ๎‚ญ๎ƒ…๎„ƒ๎ƒ๎€ƒ๎ƒฐ๎„Š๎ˆš๎€ƒ๎ƒฌ๎„ด๎„ด๎„จ๎„ฎ๎ˆš๎ˆฅ๎ˆฅ๎ƒฆ๎„๎„๎ˆ˜๎„ฎ๎„ท๎ˆฅ๎ƒฆ๎”๎ƒฆ๎ฝ๎…ก
[14] Ulker H, Kucuk D, Cantekin Z, Solmaz H. Hatay yรถresinde
๎„๎ƒ๎„ฎ๎ƒฐ๎„‰๎ƒฌ๎‚ญ๎„Š๎ƒ๎ƒŒ๎ƒ๎€ƒ๎„๎ƒ๎„ฎ๎ƒฐ๎„ƒ๎ƒ๎„Š๎€ƒ๎„ฎ๎ƒฑ๎ƒง๎ƒฑ๎„ซ๎€ƒ๎‚ญ๎„๎ƒ†๎ƒฐ๎ƒง๎ƒ๎„ซ๎„ƒ๎ƒ๎„ซ๎ƒฐ๎„Š๎ƒŒ๎ƒ๎„Š๎€ƒPasteurella multocida
ve Mannheimia haemolytica๎€ƒ๎…๎…ก๎„๎„ƒ๎‚ญ๎„ฎ๎…™๎„๎„Š๎„ท๎€ƒ๎…’๎ƒ๎€ƒ๎€„๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎…™๎„๎„ด๎ƒฐ๎„๎„ƒ๎ƒ๎„ซ๎ƒ๎€ƒ๎€ฃ๎„ท๎…™๎‚ญ๎„ซ๎„ƒ๎ƒฑ๎„ƒ๎ƒฑ๎ƒง๎ƒฑ๎€ƒ
[Isolation of Pasteurella multocida and Mannheimia haemolytica
from slaughtered bovine lung in Hatay region and detection of
๎„ด๎ƒฌ๎ƒ๎ƒฐ๎„ซ๎€ƒ๎‚ญ๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„ฎ๎„ท๎„ฎ๎ƒ†๎ƒ๎„จ๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„ƒ๎ƒฐ๎„ด๎ƒฐ๎ƒ๎„ฎ๎ˆ˜๎€ƒ๎€„๎‚“๎Ž๎€„๎€ง๎€ƒ๎€ฃ๎ƒ๎„ซ๎ƒฆ๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ ๎‡ก๎€ƒ
๎ˆซ๎ƒ†๎ƒฐ๎„ด๎ƒ๎ƒŒ๎€ƒ๎‡ ๎‡ก๎€ƒ๎Œ๎„ท๎„ƒ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ฃ๎ˆฌ๎ˆ›๎€ƒ๎‡ก๎ˆš๎‡ ๎‡Ÿ๎ˆฏ๎‡ ๎‡ฃ๎ˆ˜๎€ƒ๎€„๎…’๎‚ญ๎ƒฐ๎„ƒ๎‚ญ๎ƒ…๎„ƒ๎ƒ๎€ƒ๎ƒฐ๎„Š๎ˆš๎€ƒ๎ƒฌ๎„ด๎„ด๎„จ๎„ฎ๎ˆš๎ˆฅ๎ˆฅ๎ƒฆ๎„๎„๎ˆ˜๎„ฎ๎„ท๎ˆฅ๎…ก๎‡ฆ๎ƒฐ๎Œ๎„ซ๎Œ๎„
๎ˆซ๎‡ ๎‡ค๎ˆฌ๎€ƒ ๎€ฃ๎‚ญ๎ƒฆ๎€ƒ๎ท๎ˆ™๎€ƒ๎€บ๎„ท๎„ซ๎ƒ…๎„ท๎…ก๎€ƒ๎€„๎ˆ™๎€ƒ๎š๎…ก๎ƒ๎„Š๎€ƒ๎€พ๎ˆ™๎€ƒ๎€œ๎„ท๎…™๎„ท๎„๎€ƒ๎€น๎ˆ™๎€ƒ๎€๎ƒ๎„ƒ๎ƒ๎ƒ…๎ƒฐ๎€ƒ๎š๎ˆ™๎€ƒ๎Ž๎‚ญ๎„ซ๎‚ญ๎„‰๎‚ญ๎„Š๎€ƒ๎”๎ˆ™๎€ƒ๎€๎ƒฐ๎„ด๎ƒฐ๎„ƒ๎€ƒ๎”๎ˆ™๎€ƒ
Karakurt E. Immunohistochemical and molecular detection of
Mannheimia spp. and Pasteurella spp. in sheep with pneumonia
๎ƒฐ๎„Š๎€ƒ๎Ž๎‚ญ๎„ซ๎„ฎ๎€ƒ๎„จ๎„ซ๎„๎…’๎ƒฐ๎„Š๎ƒ†๎ƒ๎ˆฏ๎ฝ๎„ท๎„ซ๎„๎ƒ๎…™๎ˆ˜๎€ƒ๎Ž๎‚ญ๎ƒฅ๎„๎‚ญ๎„ฎ๎€ƒ๎‚€๎„Š๎ƒฐ๎…’๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ๎€น๎‚ญ๎„๎ˆ˜๎€ƒ๎€ฃ๎ƒ๎„ซ๎ƒฆ๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ
2018; 24(2):281โ€“288. doi: https://doi.org/g8wtk9
๎ˆซ๎‡ ๎‡ฅ๎ˆฌ๎€ƒ ๎‚š๎‚ญ๎„‰๎‚ญ๎„Š๎€ƒ๎ฝ๎ˆ™๎€ƒ๎€œ๎„ผ๎…™๎„ผ๎„๎ƒ…๎‚ญ๎…™๎„ซ๎‚ญ๎„‰๎€ƒ๎€พ๎ˆ™๎€ƒ๎ข๎…ก๎…™๎ƒฑ๎„ƒ๎ƒŒ๎ƒฑ๎…ก๎€ƒ๎‚ข๎ˆ™๎€ƒ๎ฝ๎ƒ๎„ซ๎…ก๎ƒฐ๎€ƒ๎€น๎ˆ™๎€ƒ๎‚€๎…™๎‚ญ๎„ซ๎€ƒ๎€„๎ˆ™๎€ƒ๎Ž๎ƒ๎„ƒ๎ƒ๎„ฎ๎€ƒ๎š๎€น๎ˆ™๎€ƒ
๎ข๎…ก๎„ฎ๎„๎…™๎€ƒ๎ท๎‚š๎ˆ™๎€ƒ๎‚š๎ƒ๎„Š๎ƒ๎„ซ๎€ƒ๎‚ข๎ˆ˜๎€ƒ๎€ฃ๎ƒ๎„ด๎ƒ๎ƒ†๎„ด๎ƒฐ๎„๎„Š๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒ…๎„๎…’๎ƒฐ๎„Š๎ƒ๎€ƒ๎„ซ๎ƒ๎„ฎ๎„จ๎ƒฐ๎„ซ๎‚ญ๎„ด๎„๎„ซ๎…™๎€ƒ๎„ฎ๎…™๎„Š๎ƒ†๎…™๎„ด๎ƒฐ๎‚ญ๎„ƒ๎€ƒ
virus, Pasteurella multocida, and Mannheimia haemolytica by
๎ƒฐ๎„‰๎„‰๎„ท๎„Š๎„๎ƒฌ๎ƒฐ๎„ฎ๎„ด๎„๎ƒ†๎ƒฌ๎ƒ๎„‰๎ƒฐ๎ƒ†๎‚ญ๎„ƒ๎€ƒ๎„‰๎ƒ๎„ด๎ƒฌ๎„๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„Š๎‚ญ๎„ด๎„ท๎„ซ๎‚ญ๎„ƒ๎„ƒ๎…™๎ˆฏ๎ƒฐ๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ƒ๎ƒŒ๎€ƒ๎ƒ†๎‚ญ๎„ด๎„ด๎„ƒ๎ƒ๎ˆ˜๎€ƒ๎Œ๎ˆ˜๎€ƒ๎‚“๎ƒ๎„ด๎ˆ˜๎€ƒ
Res. [Internet]. 2018; 62(4):439โ€“445. doi: https://doi.org/gmr4jr
๎ˆซ๎‡ ๎‡ฆ๎ˆฌ๎€ƒ ๎‚€๎ƒŒ๎ƒŒ๎ƒฐ๎„Š๎€ƒ๎ฝ๎”๎ˆ™๎€ƒ๎€๎ƒฌ๎‚ญ๎„๎„ซ๎‚ญ๎ƒ…๎„๎„ซ๎„ด๎…™๎€ƒ๎€„๎Œ๎ˆ™๎€ƒ๎Ž๎ƒฌ๎„ท๎„ฎ๎„ซ๎„๎€ƒ๎€„๎ˆ™๎€ƒ๎‚ข๎ƒฐ๎ƒŒ๎‚ญ๎„Š๎€ƒ๎€œ๎ด๎”๎ˆ™๎€ƒ๎”๎ƒฐ๎„ด๎„ซ๎‚ญ๎€ƒ๎ท๎ˆ™๎€ƒ๎€ง๎„‰๎„ซ๎‚ญ๎„Š๎€ƒ
๎ฝ๎€œ๎ˆ™๎€ƒ๎€ฃ๎ƒฌ๎‚ญ๎„‰๎‚ญ๎€ƒ๎Ž๎ˆ™๎€ƒ๎ด๎ƒฐ๎„จ๎„๎„Š๎€ƒ๎”๎Ž๎€พ๎ˆ™๎€ƒ๎€บ๎‚ญ๎ƒพ๎ƒŒ๎‚ฎ๎ƒ†๎„ฎ๎€ƒ๎”๎ˆ™๎€ƒ๎ท๎‚ญ๎ƒฌ๎ƒฐ๎ƒ…๎…ก๎‚ญ๎ƒŒ๎‚ญ๎€ƒ๎”๎‚€๎Ž๎ˆ™๎€ƒ๎€พ๎„๎„ฎ๎„ฎ๎‚ญ๎ƒฐ๎„Š๎€ƒ
๎”๎Œ๎ˆ™๎€ƒ๎Ž๎„๎ƒฐ๎„ซ๎‚ญ๎„ƒ๎‚ญ๎€ƒ๎•๎ˆ˜๎€ƒ๎€„๎„Š๎„ด๎ƒฐ๎ƒ…๎ƒฐ๎„๎„ด๎ƒฐ๎ƒ†๎€ƒ๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎ƒฐ๎„Š๎€ƒ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ƒ๎„ฎ๎ˆš๎€ƒ๎€พ๎ƒฐ๎„ฎ๎„ด๎„๎„ซ๎…™๎ˆ™๎€ƒ
mechanisms, therapeutic strategies and future prospects.
๎Œ๎ˆ˜๎€ƒ๎๎„Š๎ƒฅ๎ƒ๎ƒ†๎„ด๎ˆ˜๎€ƒ๎ฑ๎„ท๎ƒ…๎„ƒ๎ƒฐ๎ƒ†๎€ƒ๎€พ๎ƒ๎‚ญ๎„ƒ๎„ด๎ƒฌ๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ก๎‡ ๎ˆ›๎€ƒ๎‡ ๎‡ฃ๎ˆง๎‡ ๎‡ก๎ˆจ๎ˆš๎‡ ๎‡ฆ๎‡ค๎‡Ÿ๎ˆฏ๎‡ ๎‡ฆ๎‡ฅ๎‡ฅ๎ˆ˜๎€ƒ๎ƒŒ๎„๎ƒฐ๎ˆš๎€ƒ
https://doi.org/gsdt63
๎ˆซ๎‡ ๎‡ง๎ˆฌ๎€ƒ ๎”๎ƒฐ๎ƒ†๎ƒฌ๎‚ญ๎ƒ๎„ƒ๎€ƒ๎€บ๎€œ๎ˆ™๎€ƒ๎€ง๎ƒฐ๎ƒŒ๎‚ญ๎„‰๎€ƒ๎€๎ˆ™๎€ƒ๎Ž๎‚ญ๎ƒŒ๎„ƒ๎ƒ๎ƒ†๎€ƒ๎Ž๎ˆ™๎€ƒ๎”๎ƒ๎…™๎ƒ๎„ซ๎€ƒ๎Ž๎ˆ™๎€ƒ๎ท๎…“๎ƒ๎ƒ๎„Š๎ƒ๎…™๎€ƒ๎”๎ฝ๎ˆ™๎€ƒ๎”๎„ท๎„ซ๎„ซ๎‚ญ๎…™๎€ƒ
๎ด๎‚”๎ˆ™๎€ƒ๎‚”๎‚ญ๎„ด๎„ด๎„ฎ๎€ƒ๎Œ๎๎ˆ™๎€ƒ๎ท๎ƒ†๎ƒฌ๎…“๎‚ญ๎„ซ๎…ก๎€ƒ๎ท๎ˆ˜๎€ƒ๎๎„Š๎ƒ†๎„ซ๎ƒ๎‚ญ๎„ฎ๎ƒ๎ƒŒ๎€ƒ๎”๎๎€๎„ฎ๎€ƒ๎„๎ƒฅ๎€ƒ๎ƒฆ๎‚ญ๎„‰๎ƒฐ๎„ด๎ƒฌ๎„ซ๎„๎„‰๎…™๎ƒ†๎ƒฐ๎„Š๎€ƒ
and tildipirosin in the presence of the genes erm(42) and
msr(E)โ€“mph(E) for bovine Pasteurella multocida and Mannheimia
haemolytica๎ˆ˜๎€ƒ๎Œ๎ˆ˜๎€ƒ๎€„๎„Š๎„ด๎ƒฐ๎„‰๎ƒฐ๎ƒ†๎„ซ๎„๎ƒ…๎ˆ˜๎€ƒ๎€๎ƒฌ๎ƒ๎„‰๎„๎„ด๎ƒฌ๎ƒ๎„ซ๎ˆ˜๎€ƒ๎ˆซ๎๎„Š๎„ด๎ƒ๎„ซ๎„Š๎ƒ๎„ด๎ˆฌ๎ˆ˜๎€ƒ๎‡ก๎‡Ÿ๎‡ ๎‡ก๎ˆ›๎€ƒ
67(6):1555โ€“1557. doi: https://doi.org/g8wtmb
๎ˆซ๎‡ ๎‡จ๎ˆฌ๎€ƒ ๎š๎…“๎ƒ๎„Š๎€ƒ๎Œ๎ด๎ˆ™๎€ƒ๎•๎„๎…™๎ƒ๎„ฎ๎€ƒ๎•๎ˆ™๎€ƒ๎‚š๎„๎„ท๎„Š๎ƒฆ๎€ƒ๎€„๎€ง๎ˆ™๎€ƒ๎ฑ๎„ซ๎ƒฐ๎„Š๎ƒ†๎ƒ๎€ƒ๎€ฃ๎Œ๎ˆ™๎€ƒ๎€œ๎„ƒ๎‚ญ๎„Š๎ƒ†๎ƒฌ๎‚ญ๎„ซ๎ƒŒ๎€ƒ๎ฑ๎€๎ˆ™๎€ƒ
๎๎ƒ๎ƒฌ๎ƒ๎„Š๎ƒ…๎‚ญ๎„ท๎ƒ๎„ซ๎€ƒ๎ฝ๎‚”๎ˆ™๎€ƒ๎€„๎„ƒ๎…™๎€ƒ๎ท๎ท๎ˆ™๎€ƒ๎€ฃ๎‚ญ๎…’๎ƒฐ๎„ฎ๎€ƒ๎€พ๎€พ๎ˆ™๎€ƒ๎š๎ˆธ๎ด๎„๎„ท๎„ซ๎„๎ƒ๎€ƒ๎ท๎”๎ˆ™๎€ƒ๎€„๎ƒ…๎ƒŒ๎„๎€ƒ๎‚ข๎ˆ˜๎€ƒ๎‚”๎ƒฌ๎„๎„ƒ๎ƒ๎€ƒ
genome sequencing and concordance between antimicrobial
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2017;7(9):3059โ€“3071. doi: https://doi.org/ ๎ƒฌ๎„ด๎„ด๎„จ๎„ฎ๎ˆš๎ˆฅ๎ˆฅ๎ƒŒ๎„๎ƒฐ๎ˆ˜๎„๎„ซ๎ƒฆ๎ˆฅ๎…ช๎„‰๎…˜๎ƒพ๎‡ฆ
๎ˆซ๎‡ก๎‡Ÿ๎ˆฌ๎€ƒ ๎€„๎„ƒ๎ƒฌ๎‚ญ๎„‰๎‚ญ๎„‰๎ƒฐ๎€ƒ๎ฝ๎ˆ™๎€ƒ๎€๎ƒฌ๎„๎…“๎ƒŒ๎ƒฌ๎„ท๎„ซ๎…™๎€ƒ๎ฑ๎ด๎ˆ™๎€ƒ๎€บ๎„๎„‰๎ƒ๎„ฎ๎€ƒ๎•๎ˆ™๎€ƒ๎€๎‚ญ๎„ซ๎„ซ๎€ƒ๎”๎ˆ™๎€ƒ๎‚“๎ƒ๎„ƒ๎„ด๎„‰๎‚ญ๎„Š๎€ƒ๎ฝ๎ˆ™๎€ƒ๎Ž๎ƒฌ๎‚ญ๎…ก๎‚ญ๎„Š๎ƒŒ๎ƒฐ๎€ƒ
๎”๎ˆ™๎€ƒ๎”๎„๎„ƒ๎„ƒ๎ƒฐ๎„Š๎ƒฆ๎ƒ๎„ซ๎€ƒ๎Œ๎ˆ™๎€ƒ๎€ฃ๎ƒ๎„ท๎„ด๎„ฎ๎ƒ†๎ƒฌ๎ƒ๎„ซ๎€ƒ๎€„๎ฝ๎ˆ™๎€ƒ๎ฝ๎„ท๎„ซ๎„Š๎ƒฐ๎€ƒ๎€๎ˆ™๎€ƒ๎”๎‚ญ๎ƒฌ๎ƒŒ๎ƒฐ๎€ƒ๎๎ˆ˜๎€ƒ๎€น๎ƒฐ๎„ซ๎„ฎ๎„ด๎€ƒ๎ƒ๎„‰๎ƒ๎„ซ๎ƒฆ๎ƒ๎„Š๎ƒ†๎ƒ๎€ƒ๎„๎ƒฅ๎€ƒ
๎„ซ๎ƒ๎„ฎ๎ƒฐ๎„ฎ๎„ด๎‚ญ๎„Š๎ƒ†๎ƒ๎€ƒ๎„ด๎„๎€ƒ๎„‰๎‚ญ๎ƒ†๎„ซ๎„๎„ƒ๎ƒฐ๎ƒŒ๎ƒ๎„ฎ๎€ƒ๎‚ญ๎„Š๎ƒŒ๎€ƒ๎„ด๎ƒ๎„ด๎„ซ๎‚ญ๎ƒ†๎…™๎ƒ†๎„ƒ๎ƒฐ๎„Š๎ƒ๎€ƒ๎ƒฐ๎ƒŒ๎ƒ๎„Š๎„ด๎ƒฐ๎…ฅ๎ƒ๎ƒŒ๎€ƒ๎ƒฐ๎„Š๎€ƒMannheimia
haemolytica and Pasteurella multocida Isolates from beef feedlots
in Australia. Microorganisms [Internet]. 2021; 9(6):1322. doi:
๎ƒฌ๎„ด๎„ด๎„จ๎„ฎ๎ˆš๎ˆฅ๎ˆฅ๎ƒŒ๎„๎ƒฐ๎ˆ˜๎„๎„ซ๎ƒฆ๎ˆฅ๎„Š๎…˜๎…˜๎‡ฅ