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*Corresponding Author: Dereje Fufa, Assela Regional veterinary Laboratory, Assela, Oromiya, Ethiopia.
Citation: Tesfaye Belachew and Dereje Fufa, (2023), Bacterial Isolation and Identification from Bovine Mastitis and Their Pattern of Anti-Microbial Susceptibility Test in Selected Districts of North Shoa and Oromia Special Zone Surrounding Finfinne, Central Ethiopia, J. Biomedical Research and Clinical Reviews. 8(6); DOI:10.31579/2692-9406/171
Copyright: © 2023, Dereje Fufa. this is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Received: 08 December 2023 | Accepted: 15 December 2023 | Published: 22 December 2023
Keywords: mastitis; prevalence; risk factors; antimicrobial
A cross-sectional study was carried out from January 2023 to February 2023 to study the prevalence of bovine mastitis (Clinical and sub –clinical), risk factors, to isolate and identify major pathogens and to make antimicrobial susceptibility test on the isolates on cross breed lactating cows which kept under intensive farming /husbandry/ system found in Barek and Sululta town. A total of 200 lactating cows examined 66 from Sululta , 71 from Wuchale and 63 from Debre libanos for mastitis by screening test/California Mastitis Test/ for sub clinical and udder observation and palpation for clinical mastitis from 9 kebeles which found in sululta,Wuchale and Debre libanos . The overall prevalence of mastitis was 57 (28.5%), where lower in Sululta and Debrelibanos districts 16(24.24) and 15(23.80) respectively and higher in Wuchale distirict 26(36.61) by clinical and sub clinical type of mastitis. From cultured samples, the bacteria genera species and isolated were as in with high prevalence were Staphylococcus intermidius 35(71.43), Staphylococcus aureus 6(12.25),E.coli 1(2.04) .Coryne bacterium (6.12%) and Bacillus cereus 4(8.13).From currently tested few isolates using selected antimicrobial agents, all susceptible for Chloramphenicol and Gentamycin except one each isolates of S.aureus and for both which shows intermediate. Similarly most of the isolates susceptible to Erythromycin and Amoxicillin except some isolates of Staphylococcus aureus, and bacillus cereus. However, high resistance was observed by most of isolates to Penicillin and tetracycline.Those cows with different lactation stage and farming system revealed significant difference (p<0.05) in prevalence of mastitis. The prevalence of both age and parity by risk factors were insignificance (P>0.05), however, there was significance difference between semi intensive and extensive farming system (P<0.05). Prevalence of mastitis, particularly, the subclinical one could bring about major economic losses in dairy cows without notice as result of reduced milk production, poor growth or mortality of suckling calves and dam health.. The resistance to penicillin and tetracycline were observed with poor efficacy may be due to prolonged and indiscriminate usage these antimicrobials in the area so that Chloramphenicol, Gentamycin and Erythromycin could be the drugs of choice in the present study.
Mastitis is a complex and multi factorial disease, the occurrence of which depends on variables related to the animal, environment and pathogen [1]. Among the pathogens, bacterial agent is the most common one, the greatest share of which resides widely distributed in the environment of dairy cows, hence a common threat to the mammary gland [2]. Mastitis induced via pathogenic microorganisms that generally come from two sources, either environmental exposure of teat to contaminated environment, or the animal itself. Staphylococcus aureus and Streptococcus agalactiae which are predominant pathogens to cause bovine mastitis) that comprises contagious bacteria causing mastitis [3, 1].
Mastitis can be classified as clinical or subclinical depending on the presence or absence of clear clinical signs. Clinical cases of mastitis are illustrated by the presence of one or more of symptoms such as abnormal milk, udder swelling and systemic signs including elevated temperature, lethargy and anorexia [3]. Mild clinical mastitis causes flakes or clots in the milk, whereas severe cases are associated with hot, swelling and discoloration of the udder, as well as abnormal secretion. Severe clinical mastitis can also exhibit systemic reactions, such as fever and loss of appetite. The duration of infection further classifies mastitis acute or chronic manifestations, where a sudden onset defines acute cases and an inflammatory process that lasts for months and results in progressive development of fibrous tissue [4] characterizes chronic mastitis.
Mastitis can exist in the absence of visible signs of infection, and is then referred to as subclinical mastitis. It is the most prevalent form of mastitis [5]. Sub clinical mastitis are those in which no visible appearance of changes in the milk or udder, but milk production decreases, bacteria are present in the secretion and composition is altered. For every case of clinical mastitis there are 20-40 times as many cases of sub clinical mastitis [3]. The current standard method of detecting subclinical mastitis is to measure somatic cell count (SCC). Other inflammatory parameters, such as electrical conductivity, lactose, lactate dehydrogenase, acute phase proteins, etc, have been proposed as indicators of subclinical mastitis, and some have the potential of being adapted to in-line use [6].Mild cases of clinical mastitis (abnormal secretion only)may not require treatment; however, all clinical mastitis episodes accompanied by an abnormal gland or systemic signs of illness should be treated with antimicrobial agents given by intra mammary infusion (all cases) and parenterally (selected cases). Acute and peracute mastitis cases require also supportive therapy (fluid and electrolytes) and non-steroidal anti-inflammatory a gents NSAIDs [7].
The cure rate after antimicrobial treatment of clinical mastitis is very variable due to both cow and bacterial factors such as parity of the cow, chronicity of the infection and bacterial genotype [8]. To approach appropriate treatment and control measure, it is important to perform antibiotic susceptibility test on relevant and most frequently used antimicrobials. Currently, in Ethiopia, including the study area, the information on prevalence and distribution of mastitis and the sensitivity to commonly used antimicrobials for treatment of mastitis is scarce.
Improvements of the productivity of livestock sector by controlling some of the major infectious disease, has received little attention in the country, especially, mastitis, the common problem of dairies that is known by an inflammation of the mammary gland is the leading one, that can contribute to reduce, milk production [9].It is primarily resulting from an invasion of mammary tissues by pathogenic microorganisms through the teat canal resulting in physical, chemical, pathological changes in glandular tissues and milk. Evidence to date shows that affected dairy cows may loss 15% of their production and the affected quarter a 30% reduction in productivity [10]. There for the objective of the study was:
Isolations and Identifications of pathogenic bacterial from bovine mastitis and their pattern of anti-microbial susceptibility test.
2.1. Study Area
Sululta District is among one of the six districts found in Oromia special zone surrounding Addis Ababa, Oromia Regional State. It lies between 390 30‘N Latitude and 380 30' and 390 00 E longitude. It is located 40 km north west of Addis Ababa. The topography of Suluta is undulated with altitude ranging between 1600 and 3318m.a. s. l. The pattern of rainfall in the area is a bi-modal and ranges from 834 to 1440 mm. The short rainy season is between February and March while the long rainy season is between June and September. According to the Sululta Livestock Resource Development and Health office, the total cattle population of the district for the year 2011 was 210,211 heads.
Wuchale district is located at the 9º17'N-9º48'N latitude and 38º45'E-39º13'E longitude; 78 Km from Addis Ababa, the capital city of Ethiopia, and 34 Km from Fiche, the capital city of north shewa zone. The district has three major administrative towns as MukeTuri (district capital), Wobori, and Gimbichu. The highest elevation of the district is 2880m, the lowest is 1200 m, and the average elevation is 2412 m above sea level.
Debre Libanos District is located in 38o 58’ 33”E longitude and 9o 63’ 75’’N latitude with altitude ranging from 1500 to 2700 m.a.s.l. For Debre Libanos the maximum and minimum annual temperature is 23 0C and 15 0C, respectively. Its main rainy season occurs between May and September and the dry season lasts from October to April. Clay and sandy soils are the major soil types of the districts. In the district agricultural production is characterized by a mixed crop-livestock production system [11].
2.2. Study Animals
The study populations were lactating cows breeds of cattle (Holstein Friesian (HF) and Cross breed (HF x Local) in selected dairy farms in the study area. The animals were managed under Intensive, semi intensive and extensive management system. They were often provided with some supplementary diet in addition to the natural pasture and agricultural by products and some were maintained usually in separate stalls, a short distance from each other in a house. Pre-milking and post-milking hygienic procedures, such as udder washing and drying, was frequently practice.
2.3. Study design
A cross-sectional study was carried out from January to April 2023 to study the prevalence of bovine mastitis (Clinical and sub –clinical), risk factors, to isolate and identify major pathogens and to make antimicrobial susceptibility test on the isolates on cross breed lactating cows which kept under intensive farming /husbandry/ system found in Sululta, Wuchale and Debre libanos Districts.
2.4. Sampling methods and determination of sample size
Simple random sampling were carried out and the sample size of the study was determined based on sample size determination method as described by [12] with a 95% confidence interval and 5
5.1 Prevalence
A total of 200 lactating cows examined 66 from Sululta , 71 from Wuchale and 63 from Debre libanos for mastitis by screening test/California Mastitis Test/ for sub clinical and udder observation and palpation for clinical mastitis from 9 kebeles which found in sululta,Wuchale and Debre libanos (Table 1 ).The overall prevalence of mastitis was 57 (28.5%), where lower in Sululta and Debrelibanos districts 16(24.24) and 15(23.80) respectively and higher in Wuchale distirict 26(36.61) by clinical and sub clinical type of mastitis (Table 1).
Table 1- prevalence of bovine mastitis by districts
Out of 800 quarters examined during the study period 72(9.00%) were positive for mastitis while 13 (1.62%) blind (table 2).
Table 2: Quarters level examined during the study period
Those cows with different lactation stage and farming system revealed significant difference (p<0>0.05), however, there was significance difference between semi intensive and extensive farming system (P<0>
From the test results in each districts S.epidermis higher in number and E.coli was lower than all isolate as shown in the table 4.
Table 3: The prevalence of mastitis within the considered risk factors
Table 4: Summary of test result by percentage and by districts
From cultured samples, the bacteria genera species and isolated were as in with high prevalence were Staphylococcus intermidius 35(71.43), Staphylococcus aureus 6(12.25),E.coli 1(2.04) Coryne bacterium 3(6.12%) and Bacillus cereus 4(8.13) as shown in table 5 below.
Table 5: Overall prevalence of pathogen isolates causing bovine mastitis in the study area
Staphylococcus aures were highliy susceptibility (100%) to Oxacillin (OX) ,Kanamycin(K),Gentamicin(CN), Chloramphenicol (C) ,Ciprofloxacilin (Cip) Clindamycim (CD), Cefoxitin (FOX) and Vancomycin (VA) and resistant to PenicillinG(P) ,Amoxicillin(AX) and Ampicillin(AMP) as clearly explained in table 6
Table 6: Summary of Resistances and susceptibility of Staphylococcus aures bacteria by percentage
Staphylococcus epidermis were Susceptible(100) to Kanamycin (K), Gentamicin (CN) ,Streptomycin(S) ,Cefoxitin(FOX) ,Vancomycin(VA) Sulphamethaxazole-Trimethoprim(SXT25) ,Chloramphenicol(C) ,Ciprofloxacilin(Cip) and Clindamycim(CD) and resistant (100) to Amoxicillin(AX) (table 7)
Table 7: Summary of Resistances and susceptibility of Staphylococcus epidermis bacteria by percentage
E.coli was resistant to Ampicillin (AMP) and almost susceptible the other drug used for the tests as shown in table 8 below.
Table 7: Summary of Resistances and susceptibility of E.coli bacteria by percentage
The overall prevalence of mastitis in the present study was 28.50%), But this finding was lower than the 89.5% report of [13] in salale, north shoa. This could be due to difference in management system in that alternative free ranging management within door keeping system is mostly applied in the present study area. Environmental bacterial mastitis were higher in prevalence, due to poor housing facilities which predispose the accumulation of faeces on cows which will increase the rate of exposure of the teats and udder to the pathogens. Exposure to environmental Streptococci may occur during milking, between milking and during dry period [7].
From cultured samples, the bacteria genera species and isolated were as in (table 4) with high prevalence were as in (table 5) with high prevalence were Staphylococcus intermidius 35(71.43), Bacillus cereus 4(8.13), Staphylococcus aureus 6(12.25), Coryne bacterium (6.12%) and E.coli 1(2.04) from mastitis positive animal in present study shows the higher contributions of microbial in the cause of mastitis in the area.
Although Staphylococcus intermidius 35(71.43), this higher level occurrence might be due to the organism’s behavior which is adapted to survive in the udder and usually establishes a mild subclinical infection of long duration from which it shed in milk facilitating transmission to healthy animals mainly during milking [7] and the predominant pathogens isolated in this study, it was lower than the 13% reported [13]. From cultured samples, the bacteria genera species and isolated were as in with high prevalence were Staphylococcus aureus (52.62%), E.coli (10.52%) Staphylococcus intermidius., Diplocuccus spps and Staphylococcus hycus (5.26 %) and Corynebacterium bovis (3.50%) were lower than the 38.4 and 23.2% report of [13] irrespective of the agent.
From currently tested few isolates using selected antimicrobial agents, all susceptible for Chloramphenicol and Gentamycin except one each isolates of S.aureus and for both which shows intermediate. Similarly most of the isolates susceptible to erytomycine except some isolates of Staphylococcus aureus, and bacilles cereus. However, high resistance was observed by most of isolates to Penicillin and tetracycline which is agree with finding of [23] in and around Asella town
The study attempted to prevalence of bovine mastitis by considering associated risk factors in north shoa zone, isolates and determines antibiotic susceptibility profiles of pathogens involved in bovine mastitis using clinical sing, CMT, bacteriology and invitro susceptibility test. Prevalence of mastitis, particularly, the subclinical one could bring about major economic losses in dairy cows without notice as result of reduced milk production, poor growth or mortality of suckling calves and dam health. Thus, due attention should be given to increase the awareness of health management (cow-udder hygiene, milking area hygiene, milker’s hand hygiene, husbandry, tick control etc) in the area including proper antibiotic selection, dosage and frequency of administration of drugs.
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