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.

Methicillin-resistant Staphylococcus aureus isolates derived from humans and animals in Yogyakarta, Indonesia.

Background and Aim: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) as a highly pathogenic strain in veterinary and human medicine is a growing global problem. This study aimed to evaluate MRSA isolates of human and animal origin against various antibiotics in Yogyakarta, Indonesia. Materials and Methods: The susceptibility test was carried out by the disk diffusion method using Mueller-Hinton agar against nine antibiotic disks. Methicillin-resistant S. aureus strains were genetically confirmed through mecA gene detection encoding for methicillin resistance by polymerase chain reaction. Results: All 240 S. aureus strains isolated from animals and humans were resistant to penicillin G (P) (100% and 99%, respectively), followed by ampicillin (AMP), amoxicillin (AML), oxacillin (OX), erythromycin (E), clindamycin (DA), tetracycline (TE), gentamicin (GEN), and ciprofloxacin (CIP). Eighty-three MRSA strains were resistant to OX (100%), P (100%), AMP (99.27%), AML (95.52%), E (87.77%), TE (71.33%), DA (63.24%), GEN (38.81%), and CIP (26.87%). Conclusion: The antimicrobial resistance pattern of S. aureus human isolates was similar to their animal counterpart, with 77.20% of MRSA strains classified as multidrug-resistant (MDR) bacteria. These findings indicate an increase in MDR S. aureus strains of animal origin in Yogyakarta, thus raising public health concerns about MRSA zoonotic spread.

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 post-infection. 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.

Health comparison between guinea pigs raised in uncontrolled and controlled environments.

Background and Aim: Guinea pigs (GPs) (Cavia porcellus) are not only kept as pets but also widely used in biological and biomedical research. At present, GPs are also used as a species for animal-assisted therapy (AAT). Consequently, assessing their health status is vital to determining their quality of life, usability for research, and prevention of spread of potential zoonotic diseases to patients using them for AAT. GPs are mainly sourced from animal markets supplied by traditional farms, where environmental factors and sanitation are not properly controlled. This study aimed to compare health status between GPs raised in uncontrolled (conventional farm) and controlled (animal facility) environments. Materials and Methods: Sample animals were obtained from a local animal market and transported to an animal facility. After 1 week of acclimatization, the health status of the animals, including general health condition, body weight, body temperature, complete blood count, liver function (alanine aminotransferase and bilirubin), renal function (blood urea nitrogen and creatinine), and presence of ectoparasites and endoparasites, was assessed. Then, the animals were maintained in the animal facility following the standard procedure for laboratory animals. After 2 months, the animals' health status was re-examined, assessing the same parameters. Results: Based on the evaluated parameters, GPs raised in an uncontrolled environment were found to have poorer health status than those raised in a controlled environment. There were significant differences in almost all parameters between GPs raised in controlled and uncontrolled environments. We found that the populations of two ectoparasites, Gyropus ovalis and Gliricola porcelli, and one endoparasite, Eimeria caviae, decreased significantly following the movement of the animals from an uncontrolled to a controlled environment. Conclusion: GPs raised in an uncontrolled environment have poor health status. However, a controlled environment with better care management can improve the health status of GPs.

Immunosuppression by piperine as a regulator of the NLRP3 inflammasome through MAPK/NF-κB in monosodium urate-induced rat gouty arthritis.

Background and Aim: Gouty arthritis is a metabolic disorder involving monosodium urate (MSU) crystal deposition as a key initiator of acute inflammation. Dysregulation of the nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome is associated with the pathogenesis of gout through the maturation of interleukin-1ß. Piperine (PIP) is a phytochemical with an anti-inflammatory activity that has the potential as an alternative treatment for gout. In this study, we examined the effect of PIP in immunosuppression of gout inflammation through the regulation of the NLRP3 inflammasome. Materials and Methods: An in silico study was done by pharmacodynamic modeling of PIP in suppressing MSU-induced inflammation through disruption of the NLRP3 inflammasome. In vivo tests, including inflammatory assessment, histopathology, cytology, estimation of lipid peroxidation index, and detection of systemic inflammatory reactants, were performed on two groups using preventive and curative protocols. Results: In silico studies of molecular docking demonstrated the activity of PIP as a competitive inhibitor of the mitogen-activated protein kinases/nuclear factor-kappa B axis, upstream of the NLRP3 inflammasome. Analysis of gout models with curative and preventive protocols revealed the immunosuppression activity of PIP by reducing inflammatory symptoms, inhibiting tophus formation resulting from NETosis, reducing cartilage erosion, inhibiting leukocyte exudation, suppressing lipid peroxidation index, and inhibiting the production of C-reactive protein. Conclusion: The results demonstrate the activity of PIP as an immunosuppressant in gout flare. These findings indicate the potential of PIP as a candidate for prophylactic and therapeutic agent for the treatment of gouty arthritis.

Investigation of chlamydophilosis from naturally infected cats.

BACKGROUND: Chlamydophila felis, formerly known as Chlamydia psittaci var. felis, is frequently associated with ocular, respiratory, and occasionally reproduction tract infections. Even though the infection is sometimes asymptomatic, it potentially results in a latent immunosuppressive infection. OBJECTIVE: This study aimed to identify occurrences of feline chlamydophilosis, rarely reported in cats in Indonesia. METHODS: The observation was conducted in three cats with clinical signs of Cp. felis infection, particularly relapsing conjunctivitis. The cats' histories were recorded based on owners' information. Conjunctival swabs were sampled for cytology examination and molecular assay detection. A phylogenetic tree was generated using MEGA-X software to reveal group clustering. A post-mortem examination was performed on the cat that died during an examination. RESULTS: Cp. felis was detected in both cytological examination and polymerase chain reaction assay. The phylogenetic tree demonstrated that the Cp. felis isolated in this study clustered with several other isolates from the other countries. Cp. felis can be isolated from cats with different clinical manifestations and levels of severity. The chronic fatal infection demonstrated interstitial broncho-pneumonia under histopathological examination. CONCLUSIONS: Molecular assay of Cp. felis is always recommended to obtain a definitive diagnosis of feline chlamydophilosis since the disease can have various clinical manifestations. Even though it may be subclinical and is often not fatal, an infected cat may be a carrier that could spread the pathogen in the surrounding environment. Serious disease management is suggested to avoid high costs associated with regularly relapsing disease.

Potential antimicrobial properties of the Ulva lactuca extract against methicillin-resistant Staphylococcus aureus-infected wounds: A review.

Methicillin-resistant Staphylococcus aureus (MRSA), currently a major problem in hospitals worldwide, is one of the most common causes of nosocomial disease through surgical wound infection. MRSA-infected wounds have very low recovery rates and have become more problematic as some antibiotics are not effective against MRSA. Several antimicrobial and anti-inflammatory agents of green algae (Ulva lactuca) in the form of alkaloids, triterpenoids, steroids, saponins, and flavonoids have the potential to accelerate the wound healing process following MRSA wound infection. Various active compounds contained in the U. lactuca extract are thought to have multiple antibacterial and anti-inflammatory properties that can overcome the MRSA antimicrobial resistance and accelerate tissue growth in the wound healing process. This review aims to describe the potential of Ulva lactuca extract against MRSA-infected wound healing.

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.

Impact of Subclinical Haemoproteus columbae Infection on Farmed Domestic Pigeons from Central Java (Yogyakarta), Indonesia, with Special Reference to Changes in the Hemogram.

Pigeon haemoproteosis caused by Haemoproteus columbae (Apicomplexa: Haemosporida: Haemoproteidae) is globally prevalent in rock doves (Columba livia), although little is known regarding this disease in pigeons and doves in Indonesia. Blood samples of 35 farmed domestic pigeons (C. livia f. domestica) from four localities in Yogyakarta Special Region, Central Java, Indonesia, were collected from March to June, 2016, subjected to a hemogram, and analyzed for the presence of hemoprotozoan infections. Microscopic examination of blood smears revealed a prevalence of 62.5-100% of H. columbae at the four localities (n = 8-10 for each locality), and geometric means of 3.0-5.6% of erythrocytes were parasitized by young and mature gametocytes, suggesting that all infected pigeons were in the chronic phase of infection with repeated recurrences and/or reinfections. Nucleotide sequencing of mitochondrial cytochrome b gene (cytb) for haemosporidian species demonstrated the distribution of four major cytb lineages of H. columbae (mainly HAECOL1, accompanied by COLIV03, COQUI05, and CXNEA02 according to the MalAvi database). Hemogram analysis, involving the estimation of packed cell volume, erythrocyte counts, mean corpuscular volume, mean corpuscular hemoglobin concentration, and plasma protein and fibrinogen levels of 20 parasitized pigeons and five non-infected pigeons demonstrated significant macrocytic hypochromic anemia with hypoproteinemia and hyperfibrinogenemia in the infected pigeons. This study shows the profound impact of long-lasting subclinical pigeon haemoproteosis caused by H. columbae on the health of farmed domestic pigeons.

Red ginger-extract nanoemulsion modulates high blood pressure in rats by regulating angiotensin-converting enzyme production.

BACKGROUND AND AIM: Red ginger (RG) has reportedly been used in folk medicine for the management and prevention of hypertension. One of the hypertension study models in experimental animals is the unilateral ureteral obstruction (UUO). This study aimed at evaluating the effect of RG-extract (RGE) nanoemulsion on UUO-induced hypertension and angiotensin-converting enzyme (ACE) production in rats. MATERIALS AND METHODS: RG was extracted using ethanol, combined with virgin coconut oil, polysorbate 80, and polyethylene glycol 400 to form the oil phase. The particle sizes of RGE nanoemulsions were analyzed using a particle size analyzer. The UUO method was used to induce chronic kidney disease in rats (504 mg/200 g and 360 mg/200 g b/w per oral for 7 days). The systolic and diastolic blood pressure was determined non-invasively in conscious state by tail plethysmography using an automated blood pressure monitor. ACE in serum was measured using enzyme-linked immunosorbent assay. RESULTS: The RGE nanoemulsions exhibited a particle size of32.8 nm and a polydispersity index (PI) of 0.268, indicating a homogenous nanoemulsion. UUO rats treated with RGE nanoemulsion (360 mg/200 g b/w) experienced a significant decrease in both their systolic blood pressure (p<0.05) from 142±1 mmHg to 107±6 mmHg and their diastolic blood pressure from 106±1 mmHg to 84±4 mmHg. Furthermore, treatment with RGE resulted in a 10.80% decrease in the level of ACE. CONCLUSION: The size and the PI of the RGE used in this study suggest a stable and effective distribution of the particle size in the emulsions. RGE nanoemulsions at the dose of 360 mg/200 g bw can be used as potential ACE inhibitors because they were found to decrease the blood pressure of hypertensive UUO rats.

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

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.

Staphylococcus aureus Isolated from Skin from Atopic-Dermatitis Patients Produces Staphylococcal Enterotoxin Y, Which Predominantly Induces T-Cell Receptor Vα-Specific Expansion of T Cells.

While investigating the virulence traits of Staphylococcus aureus adhering to the skin of atopic-dermatitis (AD) patients, we identified a novel open reading frame (ORF) with structural similarity to a superantigen from genome sequence data of an isolate from AD skin. Concurrently, the same ORF was identified in a bovine isolate of S. aureus and designated SElY (H. K. Ono, Y. Sato'o, K. Narita, I. Naito, et al., Appl Environ Microbiol 81:7034-7040, 2015, https://doi.org/10.1128/AEM.01873-15). Recombinant SElYbov had superantigen activity in human peripheral blood mononuclear cells. It further demonstrated emetic activity in a primate animal model, and it was proposed that SElY be renamed SEY (H. K. Ono, S. Hirose, K. Narita, M. Sugiyama, et al., PLoS Pathog 15:e1007803, 2019, https://doi.org/10.1371/journal.ppat.1007803). Here, we investigated the prevalence of the sey gene in 270 human clinical isolates of various origins in Japan. Forty-two strains were positive for the sey gene, and the positive isolates were from patients with the skin diseases atopic dermatitis and impetigo/staphylococcal scalded skin syndrome (SSSS), with a detection rate of ∼17 to 22%. There were three variants of SEY (SEY1, SEY2, and SEY3), and isolates producing SEY variants formed three distinct clusters corresponding to clonal complexes (CCs) 121, 59, and 20, respectively. Most sey+ isolates produced SEY in broth culture. Unlike SEYbov, the three recombinant SEY variants exhibited stability against heat treatment. SEY predominantly activated human T cells with a particular T-cell receptor (TCR) Vα profile, a unique observation since most staphylococcal enterotoxins exert their superantigenic activities through activating T cells with specific TCR Vß profiles. SEY may act to induce localized inflammation via skin-resident T-cell activation, facilitating the pathogenesis of S. aureus infection in disrupted epithelial barriers.

Antibacterial and immunomodulator activities of virgin coconut oil (VCO) against Staphylococcus aureus.

Antibiotics have components to inhibit infections against Staphylococcus aureus, but they depend on judicious use to minimize the incidence of resistance forms. Strategies to improve the current situation include research in finding a new antimicrobial from virgin coconut oil (VCO). The saturated fatty acid, lauric acid (LA) (C12) contain in VCO was reported to have antibacterial activities. This study developed antimicrobial of VCO as an antimicrobial and immunomodulatory agent. Staphylococcus aureus used in this study had been isolated and identified from the mastitis milk crossbreed Etawa goat from Riau, Indonesia. The susceptibility of S. aureus to VCO was tested using the broth dilution method. The inhibition mechanisms of S. aureus had been studied by scanning electron microscopy (SEM) after treatment with VCO, and potential of VCO, which is known in phagocytosis macrophage. In vitro test confirmed the inhibitory effect of VCO on the growth of S. aureus at the concentration of 200 µl (equal to 0.102 % LA). Based on the result of the phagocytosing assay, VCO could increase the ability of the macrophage cells to phagocyte S. aureus significantly at a concentration of 200 µL (equal to 0.102% LA). This study concluded that the VCO could inhibit the growth of S. aureus with destructive mechanisms of bacterial cell walls and increase the ability of the phagocytic immune cells.

The Role of Sauropus androgynus (L.) Merr. Leaf Powder in the Broiler Chickens Fed a Diet Naturally Contaminated with Aflatoxin.

Aflatoxin (AF) is the secondary metabolite of Aspergillus flavus and commonly contaminates feed during storage. AF causes lowered growth rate, stress, and increased mortality in the poultry, especially for broiler industries. The aims of this study are to determine the effects of Sauropus androgynus (L.) Merr. leaf powder (SAP) in the chickens fed a diet naturally contaminated with AF. A total of 108 chickens are divided into 6 group: group I fed with basal diet (AF not detectable); group II fed with basal diet (AF not detectable) + 5% SAP; group III with AF (>1 ppb <50 ppb); group IV with AF (>1 ppb <50 ppb) + 5% SAP; group V with AF (>51 ppb <100 ppb) + 5% SAP; group VI with AF (>101 ppb <150 ppb) + 5% SAP. The data of the body weight, feed intake and efficiency, the relative weight of liver, kidney, spleen, bursa of Fabricius (BF), histopathology, haematological profile, haemagglutination inhibition (HI) titer, AF residue, and immunohistochemistry are collected on days 7, 14, and 21. All the data were analysed using SPSS 16. The supplementation of 5% SAP in the chickens fed a diet naturally contaminated with AF showed the potential effects of the body weight performance, haematological profile protection, increase in the cellular and humoral immune responses, reduction of AF residue in the organ, protection of liver, kidney, spleen, and BF histopathology, and increase in the immune-expression of CD4+/CD8+ lymphocytes ratio (P < 0.05). It shows that 5% SAP can be used as the alternative herbal supplementation to depress the impacts of aflatoxicosis in the broiler chickens.

Investigations of selected pathogens among village pigs in Central Papua, Indonesia.

Village pig husbandry is an important part of livestock production in Papua Province, Eastern Indonesia. However, high level of disease and mortality constrains production. The aim of this study was to investigate the prevalence of the selected pathogens in village pigs in the Jayawijaya Region of Papua Province, Indonesia. Two studies were conducted: Study 1 determined the prevalence of selected pathogens in dead or moribund pigs sent to the main local market for sale. Study 2 recorded the prevalence of the selected pathogens, on pig farms in the Subdistrict of Wamena that had not recorded a case of pig mortality during the duration of Study 1. Blood samples of individuals from both groups were tested for CSF antigen and antibody, as well as antibody against PCV2. Organs with evident pathological changes from Study 1 and tonsilar swabs from Study 2 were subjected to bacteriological culture and identification of Streptococcus suis and Streptococcus zooepidemicus. Faecal samples from both studies were examined for eggs of strongyle parasites, Trichuris suis, Ascaris suum, Strongyloides ransomi and coccidia. The main infections in both studies were CSF, PCV2 and strongyle parasites, but prevalence was higher in Study 1 (P < 0.05). T. suis and S. zooepidemicus were prevalent in pigs in Study 1, but rare in healthy pigs (P < 0.05). Infections with coccidia, A. suum and S. ransomi were common but did not differ between groups (P < 0.05), with S. suis infections uncommon in both studies. This suggests that infections with CSF, PCV2, strongyle and T. suis are important pathogens in village pig farms in Jayawijaya. Local pig husbandry practices, such as confining pigs and heat-treating pig feeds, may be practical solutions to help minimize infection in village pigs in Jayawijaya.

Use of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick for an easy and rapid detection of Jembrana disease virus.

Jembrana disease virus (JDV) is a viral pathogen that causes Jembrana disease in Bali cattle (Bos javanicus) with high mortality rate. An easy and rapid diagnostic method is essential for further control this disease. We used a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow dipstick (LFD), based on conserved tm subunit of Jembrana disease virus env gene. The RT-LAMP conditions were optimized by varying the concentration of MgSO4, betaine, dNTP, and temperature as well as the time and duration of reaction. The primers sensitivity for JDV was confirmed. The method was able to detect env-tm gene dilution which contained 2 × 10(-15) g of template. Comparatively, the sensitivity of RT-LAMP/LFD was 100-fold more sensitive than reverse transcription-polymerase chain reaction. The primers specificity for JDV was also confirmed using positive and negative controls. This work also showed that virus detection could be done not only on total RNA extracted from blood but various organs could also be analyzed for the presence of JDV using RT-LAMP/LFD method. The whole process, including the LAMP reaction and the LFD hybridization step only lasts approximately 75 min. Results of analysis can be easily observed with naked eyes without addition of any chemical or further analysis. The combination of RT-LAMP with LFD makes the method a more suitable diagnostic tool in conditions where sophisticated and expensive equipments are not available for field investigations on Jembrana disease in Bali cattle.

Traditional pig farming practices and productivity in the Jayawijaya region, Papua Province, Indonesia.

The objective of the current survey was to provide an update on pig farming practices in the Jayawijaya region, Papua Province, Indonesia. A structured semi-close-ended questionnaire was used to interview 367 farmers across the Jayawijaya region. Results showed that farms, on average, comprised of 8.8 pigs (CI 8.5-9.1). The average litter size was 6.0 (CI 5.7-6.3) piglets, the farrowing frequency was once a year, and the annual mortality rate was 50.2% (CI 48.4-51.9). On average, 43.4% farms (CI 36.4-50.7) allowed pigs to roam freely during daylight hours. Farmers used pigs for their own consumption (62.4%, CI 57.4-67.4), as a gift (56.6%, CI 51.5-61.7), or for sale (50.7%, CI 45.6-55.8). Veterinary services were used intensively by just 11.7% of farmers (CI 8.2-16.5). Furthermore, 34.2% (CI 29.3-39) of farmers would sell sick pigs, and 63.1% (CI 58.2-68.1) would slaughter and consume them. It was also recorded that 68.6% of farmers (CI 63.7-73.4) would eat sick pigs that had died naturally. These findings suggest that traditional pig farms in Jayawijaya are of low productivity. Moreover, the free roaming of pigs and the sale and consumption of sick pigs have the potential to allow pathogens to circulate between pig and human populations.

Genotypic characterization of Staphylococcus aureus isolated from bovines, humans, and food in Indonesia.

The present study determined the genetic relationships between 41 Staphyloccocus (S.) aureus isolates from bovines, humans, and food using a single enzyme amplified fragment length polymorphism (AFLP) technique. We evaluated the prevalence of staphylococcal enterotoxin (SE) genes and other virulence gene determinants by PCR. The identification of S. aureus was based on culturing and biochemical tests, and by amplifying a specific section of the 23S rRNA gene. PCR amplification of the SE genes (sea, seb, sec, see, seg, seh, and sei) singly or in combination was observed. Most isolates of bovine origin harbored hla (84%) and cap5 (74%), while most isolates from humans harbored hla (73%), cap8 (91%), and fnbA (100%). Strains from food sources were positive for hla (100%), cap5 (100%), and cap8 (64%) unlike isolates from humans or bovines. A single enzyme AFLP analysis revealed a correlation between AFLP clusters of some strains and the source of the isolates The genotypic results of the present study might help to better understand the distribution of prevalent S. aureus clones among humans, bovines, and food and will help control S. aureus infections in Indonesia.

Persistent occurrence of a single Streptococcus equi subsp. zooepidemicus clone in the pig and monkey population in Indonesia.

In the present study 41 mucoid growing Streptococcus equi subsp. zooepidemicus strains (37 strains isolated from healthy two from diseased pigs, two strains isolated from healthy monkeys) appeared to be phenotypically and genotypically identical to mucoid growing S. equi subsp. zooepidemicus strains isolated from a previously described outbreak among the pig and monkey population on the island of Bali, Indonesia. These findings indicate that the mucoid growing S. equi subsp. zooepidemicus clone was still present in the pig and monkey population in Indonesia.

Comparative studies on pheno- and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany.

In the present study, 35 Staphylococcal strain isolated from milk samples of 16 cows from eight farms of three different geographic locations in Central Java, Indonesia, and from milk samples of 19 cows from 19 farms of different geographic locations in Hesse, Germany, were compared pheno- and genotypically. On the basis of cultural and biochemical properties as well as by amplification of the 23S rRNA specific to Staphylococcus aureus, all isolates could be identified as S. aureus. In addition, all S. aureus isolates harboured the genes clfA and coa encoding staphylococcal clumping factor and coagulase, and the gene segments encoding the immunoglobulin G binding region and the X-region of protein A gene spa. By PCR amplification, the genes seb, seg, seh, and sei was observed for the S. aureus cultures isolated in Central Java, Indonesia and the genes sec, sed, seg, seh, sei, sej and tst for the S. aureus cultures isolated in Hesse, Germany. None of the S. aureus of both origins harboured the genes sea, see, eta and etb. All isolates were additionally positive for the genes nuc, fnbA, hla, and set1. The gene hlb was found for 6 cultures from Central Java, Indonesia and 16 cultures from Hesse, Germany. However, the gene fnbB and the gene segments cnaA and cnaB were not present among the strains isolated in Central Java, Indonesia and rare among the strains isolated in Hesse, Germany. It was of interest that most of the S. aureus isolated in Central Java, Indonesia harboured the gene cap5 and most of the strains isolated in Hesse, Germany the gene cap8. The phenotypic and genotypic results of the present study might help to understand the distribution of prevalent S. aureus clones among bovine mastitis isolates of both countries and might help to control S. aureus infections in dairy herds.