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Determination of steroid esters in hair of slaughter animals by liquid chromatography with tandem mass spectrometry
2019
Matraszek-Żuchowska, Iwona | Woźniak-Sobczak, Barbara | Sielska, Katarzyna | Posyniak, Andrzej
The use of growth promoters in animal husbandry to increase weight gain and efficiency of feed conversion into muscle has been banned in the European Union since 1988, and under Directive 96/23/EC, surveillance for anabolic steroid hormones is obligatory. The hormones present in animal tissues may be of endogenous origin or may result from illegal administration. Steps have been taken to determine selected steroids in the form of esters in the alternative matrix of animal hair. Their detection in biological material is direct proof of the illegal use of anabolics. The procedure for the determination of steroid esters in animal hair, based on digestion, extraction, purification, and liquid chromatography with tandem mass spectrometry was validated under the current regulations. In total, 348 samples of animal hair were examined using this method. Good recoveries and precision values (RSD) were obtained during validation. Decision limits (CCα) and detection capabilities (CCβ) were in the ranges of 2.57–4.18 μg kg⁻¹ and 4.38–7.12 μg kg⁻¹, respectively. The method met the criteria for confirmation techniques with respect to Commission Decision 2002/657/EC. Testing for steroid esters in animal hair was introduced into the National Residue Control Programme in 2017. Steroid esters were not found in any hair samples above the CCα, which indicates that illegal use of anabolics was not confirmed.
Show more [+] Less [-]Helpful viability of post calving tonic for the treatment of post calving complications in dairy animals
2021
Manu Kumarswamy | Vishwanath Gopal Bhagwat | Santoshkumar Tattimani | Rangesh Paramesh
Objectives: Aimless use of anti-microbials for the treatment of uterine diseases has driven the rise of safe strains. Subsequently, within the current consider, the viability of post calving tonic (PCT) was assessed in vivo for the treatment of post calving complications in dairy cattle. Materials and Methods: The placentas of 10 chosen post-calved dairy animals with a history of postpartum complications primarily held placentas were drenched with PCT 250 ml twice a day for 2 successive days. The evaluation parameters, viz., time taken for removal of lochia, placenta, involution of the uterus and estrus come back time, body condition score, and milk yield have been analyzed to assess the efficacy of PCT in post calved dairy bovines. Results: The results depicted that following the administration of PCT, the mean time taken for expulsion of uterine discharge (lochia), placenta, time taken for the involution of the uterus, and estrus come back time was 86 and 10 h, 30 and 36 days, respectively. Additionally, the administration of PCT 250 ml twice daily for 2 consecutive days to the post-calved dairy cows caused a significant increase (p < 0.05) in milk yield. Conclusion: It was apparent that PCT encourages the unconstrained removal of held placenta through its ecbolic movement and advances uterine discharge exercises. In addition, PCT supplementation caused augmentation of milk yield in post calved dairy cows. [J Adv Vet Anim Res 2021; 8(4.000): 650-655]
Show more [+] Less [-]Molecular epidemiology and antibiotic resistance pattern of Enteropathogenic Escherichia coli isolated from bovines and their handlers in Jammu, India
2014
Majueeb U Rehman | Mohd Rashid | Javeed Ahmad Sheikh | Mohd Altaf Bhat
The study was aimed to investigate the molecular epidemiology and antibiotic resistance pattern of Enteropathogenic Escherichia coli (EPEC) in bovines and their handlers in Jammu, India. A total of 173 samples comprising of 103 fecal samples from bovines (60 from cattle and 43 from buffaloes), 28 stools and 42 fingertip rinses from bovine handlers were collected during August 2011 to March 2012. The isolated 126 E. coli strains (86 from bovines and 40 from handlers) belonged to 25 different serogroups in addition to rough and untypeable strains. Using multiplex polymerase chain reaction, four EPEC strains were isolated; two each from bovines and their handlers, of which two possessed the hemolysin (hlyA) gene. The prevalence of EPEC was recorded as 1.66% (n=1/60) in cattle, 2.32% (n=1/43) in buffaloes, and 2.85% (n=2/70) in their handlers. Antibiogram studies with the EPEC revealed the presence of multi-drug resistant E. coli. The isolates were mostly resistant to Amikacin, Amoxicillin, Cefixime and Streptomycin, and sensitive to Chloramphenicol. This study indicates that bovines as well as their handlers in Jammu region harbor EPEC, many of which being multi-drug resistant and carrying the hemolysin gene could be of high pathogenic potential for humans.
Show more [+] Less [-]Detection of Y chromosome of bovine using testis specific protein and amelogenin genes
2016
Mohd Hafizal A. | Mohd Hafiz A. R. | Nor Aini W. | Suriaty R. | Halimaton Sa’adiah T. | Nurizan A.
A total of thirty-eight Mafriwal cattle were selected from a localcattle herd of a government cattle farm; of which 36 animals were sub-fertile Mafriwal female dams and two bulls which were considered as control animals (one male Mafriwal and one male Jersey). Two markers were used in the detection of Y chromosome in the sub-fertile female animal which are testis specific proteins Y-encoded (TSPY) and amelogenin (AMLX/AMLY) genes. The genes were amplified using PCR. The DNA bands from a normal male for TSPY gene size was approximately 260 bp while AMLX/ AMLY gene were approximately 341 and 467 bp. The examination of all samples showed that the sub-fertile cow revealedonly 467 bp while three fragments were detected in the control group; 260 bp (testis specific protein, Y-encoded gene), 341 and 467 bp (Amelogenin gene). The results showed that the sex chromosomeanomalies associated with Y chromosome did not occur in this group. These two sex markers can be used for the diagnosis of Y chromosome abnormality in a sub-fertile cow through polymerase chain reactionwhich is a rapid and reliable method for use in breeding herds.
Show more [+] Less [-]Macrolides-resistant Staphylococcus aureus Associated with Clinical Mastitis in Cattle and Buffalo in Egypt
2023
Khaled A. Abd El-Razik | Yousef Adel Soliman | Ehab A. Fouad | Amany A. Arafa | Soheir M. Syame | Ashraf H. Soror
Staphylococcus aureus (S. aureus), the bacteria most frequently associated with mastitis in cattle and buffalo, has a large number of genes connected to antibiotic resistance. The objective of the current research was to determine the erythromycin and macrolide resistance of S. aureus isolated from mastitic milk of cattle and buffalo, particularly resistance-related genes (ermA, ermB, ermC, ermT, and msrA). Therefore, 150 dairy cattle and 135 dairy buffalo bred by small farmers in various governorates of Egypt (Cairo, Giza, Kalyobia, Fayoum, and Kafr El-Sheikh) provided a total of 285 milk samples. Inspection revealed that a total of 34 (22.7%) and 36 (26.7%) milk samples from cattle and buffalo, respectively, had clinical mastitis. With a total recovery of 31 (44.3%) S. aureus isolates. Bacterial isolation and identification of S. aureus verified the isolation and identification of 14 (41.2%) and 17 (47.2%) S. aureus isolates from cattle and buffalo, respectively. Utilizing a TaqMan probe-based real-time PCR method that targets the nuc gene, all S. aureus isolates were verified. In instances of bovine mastitis in India and Kenya, conventional PCR targeting the nuc gene, followed by DNA sequencing and phylogenetic analysis, revealed a high homology (100%) with that of S. aureus strains isolated from milk. For the tested genes, the prevalence of resistant strains was 9.6% (ermA), 64.5% (ermB), 70.9% (ermC), 19.3% (ermT), and 9.6%. (msrA). Therefore, effective control measures should be adopted to stop the spread of drug-resistant S. aureus to humans.
Show more [+] Less [-]Preliminary study on classification of raw bovine’s milk using ATR-FTIR coupled with PCA from Peninsular Malaysia
2017
Ketty G. S. L. | Norakmar I. | Falizah I. | Kamaliah G. | Izwan I. | Faridah I. | Khairunnisak M. | Saipul B. A. R. | Marni S. | Roosnoor F. H.
A classification of raw bovine’s milk samples according to theirgeographical origin in Peninsular Malaysia by Makmal Kesihatan Awam Veterinar, Department of Veterinary Services Malaysia (DVS). Six hundred bovine milk samples were collected from Perlis, Kedah, Perak, Selangor, Pahang, Negeri Sembilan, Melaka and Johor states by 26 milkcollecting centres under DVS. This study was carried out directly using attenuated total reflectance Fourier transform infrared(ATR-FTIR) spectroscopy method coupled with a multivariate principal component analysis (PCA). The spectra generated by ATR-FTIR were analysed and regions of interest were found in between 3851.651cm-1 until 2700.819 cm-1 and 2419.173 cm-1 until 977.368 cm-1. The absorbance and wavenumber data of the regions were then analysed using PCA and the results show presence of clustering towards theirgeographical origin. ATR-FTIR coupled with multivariate PCA has potential for classifying the geographical origin of raw milk produced within Peninsular Malaysia. This method provides a rapid and nondestructive secondary methodology in milk classification without further sample preparation.
Show more [+] Less [-]Molecular prevalence and species co-infection of bovine haemoparasites in Peninsular Malaysia
2017
Ibrahim A. R. | Maizatul A. M. | Chandrawathani P. | Jesse, F. F. A. | S. D., Ola-Fadunsin | Sani, R. A. | Amlizawathy A. | Sharma, R. S. K.
Bovine haemoparasites are cosmopolitan in distribution and are known to cause substantial losses to the cattle industry. In spite of their economic importance, there remains a dearth of information on their molecular epidemiology in many parts of the world including Malaysia. To ascertain the molecular prevalence and species co-infection of bovine haemoparasites in the country, blood samples were collected from 1,045 heads of beef and dairy cattle on 43 farms from six geographical zones throughout Peninsular Malaysia. Samples subjected to PCR amplification of parasite species-specific genetic fragments revealed that Anaplasma marginale was the most prevalent haemoparasite (72.6%),followed by Theileria orientalis(49.8%),Candidatus Mycoplasma haemobos ( 47. 0 % ),Babesia bovis(32. 5%), Babesia bigemina (30.5%) and Trypanosomaevansi(17.9%). A high percentage (92.1%) of cattle was infected with either one or more haemoparasites. Triple haemoparasite species co-infection was the most prevalent (25.6%), followed closely by double species co-infection (25.1%). The most common (8.8%) and significantly correlated(rs= 0.250; p<0.01) combination was A. marginale+ T.orientalis. The present study constitutes the first attempt in the country to document the molecular prevalence and species co-infection of bovine haemoparasites over a wide spatial distribution. The data obtained will facilitate treatment, control and prevention measures to improve the local cattle industry.
Show more [+] Less [-]Isolation and molecular characterization of Brucella abortus and Brucella melitensis from samples received by the Regional Veterinary Laboratory, Bukit Tengah, Malaysia
2017
Thenamutha M. | Zakiah M. D. | Azizul O. | Maswati M. A.
A study was carried out to report the phylogenetic analysis of Brucella abortus and Brucella melitensisby using molecular techniques from samples submitted to the Regional Veterinary Laboratory, Bukit Tengah.In this study, identification and genetic characterization of Brucella isolated samples using molecular analysis based on IS711 sequence between localisolates and foreign countries accesses in GenBank was done successfully. A total of 31 samples were isolated for Brucella species and then were amplified byPCR, directly sequenced and compared genetically to published sequences which were obtained from GenBank. The most common Brucella species that was found in both bovine (76.5%) and caprine (85.7%) through diagnostic samples in Regional Veterinary Laboratory, Bukit Tengah, was Brucella melitensis. PCR and sequencing were confirmed positive with 76.5% for Brucella melitensis, 23.5% for Brucella abortus and 23.5% for mixed infectionfrom the total of 17 bovine samples. In caprine, the detection of Brucella melintesis and Brucella abortus showed 85.7% and 21.4% respectively meanwhile total mixedinfection showed 21.4%. These clustering between local isolates of Brucella melitensis were phylogenetically related to other Asian countries such as Singapore,Yemen and Saudi Arabia. The Neighbour Joining Analysis clustered the Brucella abortus local isolates for both bovine and caprine were most closely related to India,Iran, Italy and USA. Interestingly, all the isolates within Malaysia have a close relationship (>95%) with the low level of genetic diversity. When local isolates arecompared to GenBank data, it gives an indication on the possible sources of these infections. Eventually, it will improve the import and export policies to controlbrucellosis in Malaysia.
Show more [+] Less [-]Bovine intestinal cellular responses following primary and challenge infections with Calicophoron microbothrium metacercariae
2008
Mavenyengwa, M.(University of Zimbabwe Department of Paraclinical Veterinary Studies Faculty of Veterinary Science) | Mukaratirwa, S.(University of KwaZulu-Natal School of Biological and Conservation Sciences) | Obwolo, M.(University of Zimbabwe Department of Paraclinical Veterinary Studies Faculty of Veterinary Science) | Monrad, J.(Danish Centre for Experimental Parasitology)
This study was carried out to establish whether cattle can develop resistance to re-infection by Calicophoron microbothrium by assessing the response of intestinal mucosal globule leukocytes, eosinophils, mast cells and basophils, and the establishment of the parasite in the host. A total of 24 1-year-old Tuli steers were randomly divided into four groups of six animals each and infected with C. microbothrium metacercariae. On the first day of the study, animals in Groups I and II were immunized with 5 000 metacercariae and then challenged with 15 000 metacercariae on Day 150 post-immunization. Animals in Group III were immunized with 15 000 metacercariae at the same time that Groups I and II animals were challenged to act as a positive control group. Animals in Group IV were left uninfected and acted as a negative control group. Three animals from each group were slaughtered on Day 28 post-challenge and the remainder were slaughtered on Day 42 post-challenge. The established amphistomes were recovered and histopathological and cytological examinations were done on the jejunum, duodenum, abomasum and the rumen. The establishment rates of the challenge infection in the immunized and challenged groups were lower and ranged from 0 to 0.2 % as compared to 6 %> from naive animals infected as positive controls. Animals immunized and then challenged with C. microbothrium had significantly higher eosinophil, mast cell and globule leukocytes counts in the intestinal mucosa (P < 0.05) as compared to those of the control group. The study indicates that cattle can develop resistance to C. microbothrium re-infection and that eosinophils and mast cells may be important cells in the rejection of the parasite.
Show more [+] Less [-]Enzyme-linked immunosorbent assay (ELISA) as a diagnostic tool for Guatemalan onchocerciasis using a bovine filaria (Onchocerca gutturosa) antigen and blood samples collected on filter paper
1983
Ito, M. | Lujan-T, A. (Servicio Nacional de la Erradicacion de la Malaria, Ministerio de Salud Publica (Guatemala)) | Fukumoto, S. | Kamiya, M.