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Development of ELISA against milk haptoglobin for diagnosis of subclinical mastitis in goats.

ABSTRACT: The study described the development of a haptoglobin-based diagnostic tool for mastitis in Ettawa crossbreed goats. Fifty eight milk samples were collected from a flock of goats in Yogyakarta, Central Java, Indonesia. All samples were tested for mastitis using the California Mastitis Test (CMT), Somatic Cell Count (SCC), and Polymerase Chain Reaction (PCR) to identify Staphylococcus aureus, Streptococcus uberis and Streptococcus agalactiae. The presence of haptoglobin mRNA and proteins in the milk somatic cells was detected using Sanger sequencing and SDS-PAGE, respectively. Milk haptoglobin levels were subsequently estimated using an indirect enzyme-linked immunosorbent assay (ELISA) developed in this study. The Receiver Operating Characteristic (ROC) analysis was performed to compare the sensitivity and specificity of CMT, SCC, and the ELISA using the PCR as the reference standard. Kappa test was used to determine the agreement between the three imperfect tests. Results indicated that somatic cells of goat milk expressed a haptoglobin mRNA with a size of 174 bp and two haptoglobin proteins with molecular weights of 18 kDa and 32 kDa. The PCR test showed that 81% of samples were diagnosed positive for mastitis. At a specificity level of 50%, the ROC indicated that the ELISA was more sensitive compared to SCC or CMT (consecutively, 96%, 94%, and 92%). Kappa values between haptoglobin ELISA and CMT or SCC were high (0.84 and 0.81, respectively). This study indicates that somatic cells of goat milk were capable of synthesizing and secreting haptoglobin. Milk haptoglobin can be a potential target for an early detection of mastitis in goats.

SUBMITTER: Windria S

PROVIDER: S-EPMC7907475 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Development of ELISA against milk haptoglobin for diagnosis of subclinical mastitis in goats.

Windria Sarasati S Salasia Siti Isrina Oktavia SIO Nugroho Widi W Widayanti Rini R Indarjulianto Soedarmanto S

Heliyon 20210222 2

The study described the development of a haptoglobin-based diagnostic tool for mastitis in Ettawa crossbreed goats. Fifty eight milk samples were collected from a flock of goats in Yogyakarta, Central Java, Indonesia. All samples were tested for mastitis using the California Mastitis Test (CMT), Somatic Cell Count (SCC), and Polymerase Chain Reaction (PCR) to identify Staphylococcus aureus, Streptococcus uberis and Streptococcus agalactiae. The presence of haptoglobin mRNA a ...[more]

PMID: 33665459

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Project description:A cross-sectional study was carried out to determine the prevalence and risk factors of subclinical mastitis in dairy goats in Thika East Subcounty, Kenya. Further the bacterial pathogens and their antibiogram were investigated. Farm level data on risk factors were obtained from 41 farmers using questionnaires. Milk was obtained from 110 lactating dairy goats and tested for submastitis using California Mastitis Test (CMT). The prevalence of subclinical mastitis at goat level was estimated to be at 50.9% using CMT, out of which 86.5% yielded bacteria on culture. The significant risk factors associated with the occurrence of subclinical mastitis were cleaning schedule (p=0.022, OD=1.047) and parity of the goat (p=0048, OD=1.37). Higher prevalence of subclinical mastitis was observed for goats residing in houses cleaned at least once a fortnight. Does in the first parity were least affected. 169 bacterial isolates were obtained from culture, of which 52 isolates from major classes of isolated bacteria were tested for antibiotic sensitivity to six antibiotics. Fourteen different bacteria were isolated and identified from the milk samples. Coagulase-negative Staphylococci (20.7%), Serratia spp. (19.5%), Citrobacter spp. (16%), Klebsiella spp. (11%), Staphylococcus aureus (10.7%), Enterobacter spp. (6.5%), Escherichia coli (5.9%), Proteus spp. (3%), Corynebacterium spp. (1.8%), Morganella spp. (1.8%), Streptococcus spp. (1.2%), Providencia spp. (0.6%), Micrococcus spp. (0.6%), and Staphylococcus intermedius (0.6%) were isolated and identified from the samples. All the isolates were resistant to Penicillin G, while 98% of the isolates were sensitive to Streptomycin. In conclusion, the study showed that a large proportion of goats were affected by subclinical mastitis, with the main bacteria being Staphylococci spp. and coliforms, and that most of the tested antibiotics can be used in the treatment of mastitis. Farmers need to be trained on improved control of mastitis through adoption of good dairy husbandry and milking practices.

Dataset Information

Development of ELISA against milk haptoglobin for diagnosis of subclinical mastitis in goats.

Publications

Development of ELISA against milk haptoglobin for diagnosis of subclinical mastitis in goats.

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