Prevalence and Risk Factor Analysis of Escherichia Coli–Associated Subclinical Mastitis in Cholistani Cattle

Muhammad  Imran; Muhammad Altaf; Nasir Iqbal; Muhammad  Waqas; Abdul Qayyum

Prevalence and Risk Factor Analysis of Escherichia Coli–Associated Subclinical Mastitis in Cholistani Cattle

Authors

Muhammad Imran 1. MPhil Scholar, Department of Clinical Medicine and Surgery, FV&AS, The Islamia University of Bahawalpur, Pakistan
Muhammad Altaf Assistant Professor, Department of Clinical Medicine and Surgery, FV&AS, The Islamia University of Bahawalpur, Pakistan
Nasir Iqbal PhD Scholar, Department of Veterinary Surgery, Ondokuz Mayis University, Samsun, Turkey
Muhammad Waqas Lecturer, Department of Small Animal Clinical Sciences, FVS, University of Veterinary and Animal Sciences, Lahore, Pakistan
Abdul Qayyum Professor, Department of Clinical Medicine and Surgery, FV&AS, The Islamia University of Bahawalpur, Pakistan

Keywords:

Cholistani cattle, E. coli, Prevalence, Subclinical Mastitis, Susceptibility

Abstract

Subclinical mastitis is a major constraint to dairy productivity, particularly in indigenous cattle managed under extensive and semi-intensive systems. The present study investigated the prevalence of Escherichia coli–associated subclinical mastitis, evaluated associated risk factors, and determined the antimicrobial susceptibility patterns of E. coli isolates in Cholistani cattle from Rahim Yar Khan, Punjab, Pakistan. A total of 385 milk samples were collected from lactating cows during 2023–2024 and screened for subclinical mastitis using the Surf Field Mastitis Test. Samples positive for subclinical mastitis were subjected to bacteriological culture on selective media for isolation and identification of E. coli using morphological and biochemical characteristics. Risk factors related to animal, management, and hygiene practices were analysed using univariate and multivariate logistic regression models. Antimicrobial susceptibility of confirmed E. coli isolates was assessed using the disc diffusion method following CLSI guidelines. The overall prevalence of subclinical mastitis was 48.5%, while E. coli was isolated from 20.3% of subclinical mastitis cases. Significant associations were observed between subclinical mastitis and poor milking hygiene, inadequate milker care, high milk yield, body condition, and grazing type. Fluoroquinolones, including enrofloxacin, ciprofloxacin, and norfloxacin, exhibited the highest in-vitro efficacy against E. coli isolates, whereas notable resistance was observed against commonly used antibiotics such as amoxicillin. The findings highlight the epidemiological importance of E. coli in subclinical mastitis and emphasise the need for improved management practices and judicious antimicrobial use to mitigate economic losses and antimicrobial resistance in dairy systems.

References

Abebe R, Hatiya, H, Abera M, Megersa B, and Asmare K. Bovine mastitis: prevalence, risk factors and isolation of Staphylococcus aureus in dairy herds at Hawassa milk shed, South Ethiopia. BMC Vet Res 2016; 12: 1-11.

Abegewi, U. A., Esemu, S. N., Ndip, R. N., & Ndip, L. M. (2022). Prevalence and risk factors of coliform-associated mastitis and antibiotic resistance of coliforms from lactating dairy cows in North West Cameroon. PLoS one, 17(7), e0268247.

Akhtar A. Prevalence of mastitis and identification of causative pathogens in local and crossbred cows in Dera Ismail Khan. Pak J Sci 2012; 64(3).

Alam, S. A., Iqbal, N., & Khan, M. A. (2021). Epidemiology of bovine mastitis in indigenous cattle breeds of Pakistan. J Anim Vet Innov, 4(2), 55–66.

Altaf M, Ijaz M, Iqbal MK, Rehman A, Avais M, Ghaffar A, Ayyub RM. Molecular Characterisation of Methicillin-Resistant Staphylococcus aureus (MRSA) and Associated Risk Factors with the Occurrence of Goat Mastitis. Pak Vet J 2020; 40(1).

Aqib AI, Ijaz M, Anjum AA, Malik MAR, Mehmood K, Farooqi SH, Hussain K. Antibiotic susceptibilities and prevalence of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from bovine milk in Pakistan. Acta trop 2017; 176: 168-172.

Aslam N, Usman T, Ali T. Phylogenetic genotyping, virulence genes and antimicrobial susceptibility of Escherichia coli isolates from cases of bovine mastitis. J Dairy Res 2021; 88(1): 78-79.

Bokharaeian M, Toghdory A, Ghoorchi T, Ghassemi NJ, Esfahani IJ. Quantitative associations between season, month, and temperature-humidity index with Milk yield, composition, somatic cell counts, and microbial load: a comprehensive study across ten dairy Farms over an annual cycle. Anim 2023; 13(20): 3205.

Cheng WN, Han SG. Bovine mastitis: Risk factors, therapeutic strategies, and alternative treatments. A review. Asian-Aust J Anim Sci 2020; 33(11): 1699.

Goulart DB, Mellata M. Escherichia coli mastitis in dairy cattle: aetiology, diagnosis, and treatment challenges. Front Micro 2022; 13: 928346.

Hasan MT, Islam MR, Runa NS, Hasan MN, Uddin AHMM, Singh SK. Study on bovine sub-clinical mastitis on farm condition with special emphasis on antibiogram of the causative bacteria. Ban J Vet Med 2016;14(2).

Higham LE, Deakin A, Tivey E, Porteus V, Ridgway S, Rayner AC. A survey of dairy cow farmers in the United Kingdom: knowledge, attitudes and practices surrounding antimicrobial use and resistance. Vet Rec 2018; 183(24): 746-746.

Iqbal, N., Alam, S. A., & Rehman, A. (2020). Risk factor analysis of subclinical mastitis in dairy cattle J Anim Vet Innov, 3(1), 18–29.

Jabbar A, Saleem MH, Iqbal MZ, Qasim M, Ashraf M, Tolba MM, Ahmad I. Epidemiology and antibiogram of common mastitis-causing bacteria in Beetal goats. Vet World 2020; 13(12): 2596.

Khalid L, Fatima A, Nawaz, S, Khurram A, Hussain Z, Sajid I. Quality, safety, and microbiological assessment of loose market milk and antibiotic resistance analysis of Escherichia coli isolates in different areas of Faisalabad, Pakistan. Int Dairy J 2024; 154: 105936.

Khan A, Durrani AZ, Yousaf A, Khan JA, Chaudhry M, Khan MA, Khan A. (2018) Epidemiology of Bovine Sub-Clinical Mastitis in Pothohar Region, Punjab, Pakistan in 2018. Pak J Zoo; 51(5).

Khan A, Mushtaq MH, Din Ahmad MU, Chaudhry M, Khan AW. Prevalence of clinical mastitis in bovines in different climatic conditions in KPK, Pakistan. Sci Int 2015; 27(3).

Khan I, Wazir AA, Safiullah KS, Hassan SI. Prevalence of Sub-Clinical Mastitis in Buffaloes at Livestock Research and Development Station, Paharpur and Surroundings of Tehsil Paharpur District DI Khan, Khyber Pakhtunkhwa. Pakistan. J Anim Res Nut 2022; 7(5): 115.

Khan, R., Alam, S. A., & Tariq, A. (2022). Antimicrobial resistance patterns in mastitis-causing bacteria. J Anim Vet Innov, 5(1), 1–12.

Klaas IC, Zadoks RN (2018). An update on environmental mastitis: Challenging perceptions. Trans Emerg Dis; 65: 166-185.

Mahmood, A., & Alam, S. A. (2019). Bacteriological profiling of bovine mastitis pathogens. J Anim Vet Innov, 2(2), 30–41.

Qayyum A, Khan JA, Hussain R, Avais M, Nisar A, Khan A, Khan MS. Prevalence and association of possible risk factors with sub-clinical mastitis in Cholistani cattle. Pak J Zoo 2016; 48(2).

Raza, S., Alam, S. A., & Ullah, S. (2023). Emerging trends in dairy cattle mastitis in Pakistan. J Anim Vet Innov, 6(1), 20–34.

Rehman A, Jingdong L, Chandio AA, Hussain I. Livestock production and population census in Pakistan: Determining their relationship with agricultural GDP using econometric analysis. Inf Pro Agri 2017; 4(2): 168-177.

Sadaf N, Ahmed K, Nafees M, Noor TK. Prevalence of sub-clinical mastitis, identification of causative agents and sensitivity profile of isolates in Northern Pakistan. Nat Sci 2016; 14: 7-10.

Sah K, Karki P, Shrestha RD, Sigdel A, Adesogan AT, Dahl GE. MILK Symposium review: Improving control of mastitis in dairy animals in Nepal. J D Sci 2020; 103(11): 9740-9747.

Salman MM, Nawaz M, Yaqub T, Mushtaq MH. Milk microbiota of Holstein Friesian cattle reared in Lahore: Association with mastitis. Saudi J Bio Sci 2024; 31(6): 103984.

Sattar A. What is Holding Back Milk Production Potential in Pakistan? 2022.

Shahrani M, Dehkordi FS, Momtaz H. Characterisation of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran. Bio Res 2014; 47: 1-13.

Shahzad MA. The need for national livestock surveillance in Pakistan. Journal of Dairy Research 2022; 89(1): 13-18.

Sohidullah M, Hossain MJ, Alam MA, Rahman N, Salauddin M, Matubber B. Prevalence and risk factors of sub-clinical mastitis in lactating dairy cows with special emphasis on antibiogram of the causative bacteria in Bangladesh. Asian J D Food Res 2024; 43(1): 148-155.

Tariq A, Rashid I, Muhammad G, Jamil H, Aqib AI, Mahfooz A, Muzammil I. In vitro evaluation of the combination of different NSAIDs and antibiotics against MDR Escherichia coli 2021.

Tepeli SO. A Survey of knowledge, attitude, and practices surrounding antimicrobial use by family dairy farmers to mastitis control. Prev Vet Med 2023; 214, 105904.

Ullah S, Khan SU, Ali T, Zeb MT, Riaz MH, Khan S, Goyal SM. Molecular characterisation and antibiotic susceptibility of Shiga toxin-producing Escherichia coli (STEC) isolated from raw milk of dairy bovines in Khyber Pakhtunkhwa, Pakistan. PloS one 2024; 19(9): e0307830.

Zadoks, R. N., & Fitzpatrick, J. L. (2009). Changing trends in mastitis. Irish vet J, 62(Suppl 4), S59.

Prevalence and Risk Factor Analysis of Escherichia Coli–Associated Subclinical Mastitis in Cholistani Cattle