The Empowerment of Livestock Farmers in Subclinical Mastitis Test with GAMA Anti-Haptoglobin in “Sahabat Ternak” Etawah Crossbreed Goat Farm

Sahabat Ternak is one of the Etawah crossbreed (PE) goat farm groups in Sleman. This farm group focuses on goat milk production and processed goods. Problems that commonly arise in dairy goat farms are the cases of subclinical mastitis, which are quite high. This disease may cause a decrease in milk production and quality. The mastitis subclinical detection method which is often used by the farming community is the somatic cell count (SCC) and California mastitis test (CMT). However, both tests have low accuracy. Recently, a new method named GAMA Anti-Haptoglobin, which is more accurate and can be done easily has been established by livestock farmers. This community service aims to empower livestock farmers in applying GAMA Anti-Haptoglobin as a sensitive, rapid, and accurate subclinical mastitis detection kit in Sahabat Ternak goat farm. The method used in this activity consisted of discussion, socialization, and training for livestock farmers, as well as laboratory testing, evaluation of test findings, and treatment for PE goats. After the training, the livestock farmers were able to apply GAMA Anti-Haptoglobin mastitis detection method effectively. The implementation of this easy and accurate field mastitis detection method, as well as personnel with reliable skills, will support in the decrease of mastitis cases and increase milk production and quality, as well as community welfare

Development of antibodies against recombinant staphylococcal enterotoxin B from food poisoning cases

Background and Aim: Staphylococcal enterotoxin B (SEB) is the most common serotype involved in food poisoning. The aim of this study was to develop immunoassay detection methods using a recombinant enterotoxin B antigen protein to produce recombinant polyclonal antibodies in vivo. Materials and Methods: Staphylococcus aureus isolated from a food poisoning case (strain JH5800) was analyzed by polymerase chain reaction (PCR) and confirmed to contain a seb gene of 477 bp. A SEB segment was amplified, cloned, sequenced, and aligned. The PCR product corresponding to the predicted mature SEB peptide was inserted into Escherichia coli BL21 (DE-3) expression vector and expressed as a hexahistidine-SEB fusion protein. Antiserum against recombinant SEB protein was produced by immunization of Balb/c mice. Results: In the indirect enzyme-linked immunosorbent assay (ELISA), the polyclonal antibodies produced had a titer of 1:3200. The seb gene of Staphylococcus aureus isolated from a poisoning case (JH5800) had a molecular size of about 477 bp and a band of recombinant SEB toxin was observed at approximately 30 kDa on SDS-PAGE gel. The polyclonal anti-SEB antibody titer, as revealed by indirect ELISA, was 1:3200 at 59 days. Conclusion: SEB recombinant protein could be used to produce polyclonal antibodies. ELISA and Western blotting were used to analyze the specificity and sensitivity of the recombinant polyclonal antibodies. Polyclonal antibodies produced could be used to detect SEB on a large-scale

Distribution of antimicrobial resistance genes of methicillin-resistant Staphylococcus aureus isolated from animals and humans in Yogyakarta Indonesia

Background and Aim: Methicillin-resistant Staphylococcus aureus (MRSA) has been known as a highly pathogenic bacteria in animals and humans, which is still becoming a global health issue. The prevalence of MRSA infection continues to increase worldwide and has become a global concern as a dangerous zoonotic disease. The World Health Organization estimates that by 2050 MRSA will be the leading cause of death. This study aimed to estimate the prevalence of MRSA in S. aureus isolates of veterinary and human origin in Yogyakarta, Indonesia. Materials and Methods: A total of 42 cases of S. aureus infection were examined in this study, consisting of nine isolates from cattle, five from goat, and 28 from human. All isolates were confirmed as S. aureus based on bacterial culture and detection of 23S rRNA and thermonuclease nuc gene by polymerase chain reaction (PCR). Results: Among 42 isolates, 35 isolates (83.3%) were identified as MRSA by PCR positive of mecA gene encoding methicillin resistance. Most MRSA strains were found in human isolates (100%), followed by cattle isolates (55.5%) and goats (40%). All MRSA strains were also multi-resistant to penicillin (blaZ gene) and tetracycline (tetK, and tetM genes) with a prevalence of about 98%. Conclusion: MRSA prevalence in humans and animals has increased significantly in Yogyakarta, Indonesia, compared to the previous study. The antimicrobial resistance pattern of MRSA animal isolates tends to be similar to humans and, thus, raises public health concerns about MRSA zoonotic spread

Streptococcus equi subsp. zooepidemicus finding in confirmed feline infectious peritonitis cat patient

Background: Feline infectious peritonitis (FIP) is a fatal immune-mediated disease in cat, caused by mutated feline coronavirus (FCoV). Due to its difficulties in diagnosis, FIP is sometimes underdiagnosed. Therefore, several laboratory procedures were performed to gain high index suspicion of FIP. However, through several laboratory findings, not only FIP but also SEZ infection was confirmed in this case. Case description: A-year-old male, domestic cat was admitted to Veterinary Medicine Clinical Pathology Laboratory, Universitas Gadjah Mada, for further effusion examination due to its high suspicion of feline infectious peritonitis (FIP). Further examination using molecular and post-mortem analysis resulted on confirmed SEZ infection and FIP. This study informed the manifestation and pathological changes in patient with SEZ and FIP in the same time. Conclusions: This study showed that viral infection followed by bacterial infection could be fatal and untreatable. After these findings, clinicians may consider SEZ infection in cat with respiratory disorder followed by thoracic effusion besides FIP. Companion animal, especially outdoor-kept animal, possibly become infected from its contact to another human or animal in the environment

Cellular immune response of Staphylococcus aureus enterotoxin B in Balb/c mice through intranasal infection

Background and Aim: Staphylococcus aureus produces various superantigen exotoxins, including staphylococcal enterotoxin B (SEB). It causes fatal anaphylactic reactions and toxic shock. This study aimed to evaluate the reaction of leukocytes and histopathological changes in the respiratory organs of Balb/c mice after intranasal infection with enterotoxigenic S. aureus (SEB). Materials and Methods: The presence of the seb gene in S. aureus was established in this study using polymerase chain reaction-specific primer. Two groups of 8-week-old male Balb-c mice consist of six mice in each group. The treated group was infected with 50 μL and 100 μL of SEB intranasal on days 1 and 14, respectively. NaCl was administered in the second group and was considered as a control group. Blood samples were collected through the retro-orbital plexus on days 1, 4, 7, 14, and 22 after infections. Total cell counts were analyzed with an independent sample t-test and compared using the statistical package for the social sciences (SPSS) version 16.0 (IBM Corp., NY, USA). The infected tissues of the respiratory organ were observed descriptively and compared to the control group. Results: The seb gene with a molecular size of 478 bp, indicating the SEB strain, is present in S. aureus used in this study. Intranasal administration of SEB showed increased leukocytes, lymphocytes, monocytes, and eosinophils on day 22 postinfection. Significant leukocytosis was seen on days 6 and 14; lymphocytosis on days 1, 4, 6, and 16; and eosinophilia on days 6, 14, and 22 compared with the control group (p > 0.05). In contrast, the neutrophil decreased after an increase of immature band cells compared to the control group, indicating a severe acute infection with SEB. The lungs and trachea of the test group had an inflammatory cell accumulation in the respiratory organ. Conclusion: Intranasal route infection of S. aureus containing seb gene significantly induced the cellular immune response and caused pathological changes in the respiratory tissues of the Balb/c mice model. The hematological changes were aligned with marked pathological changes in the respiratory tract. Balb/c mice could be an excellent experimental model to study toxic and anaphylactic shock against SEB to define the future therapeutic agents