TRYPANOSOMA EVANSI AND NEOSPORA CANINUM AMONG WATER BUFFALOES (BUBALUS BUBALIS) IN THE PHILIPPINES

The study determined the positivity rate of Trypanosoma evansi and Neospora caninum antibodies in water buffaloes in the province of Nueva Ecija, Philippines using Polymerase Chain Reaction (PCR) for T. evansi and competitive Enzyme-linked Immunosorbent Assay (cELISA) for N. caninum antibodies . A total of 100 whole blood and 100 serum samples were collected to test for T. evansi and N. caninum , respectively. Rotat 1.2 VSG gene was target using PCR for T. evansi detection. Neospora caninum antibody detection was done from the serum samples using cELISA test kit.Results revealed that the positivity rate of T. evansi in Nueva Ecija was 11% (11/100). The positive animals identified were from the municipalities of Muñoz (4/16; 25%), Sta. Rosa (3/13; 23.08%) and Talugtug (4/16; 25%). The seropositive rate of Nueva Ecija for N. caninum. was 46% (46/100), seropositive animals were identified in Cabanatuan City, 57.14% (4/7); Science City of Muñoz, 43.14% (22/51); Sta. Rosa, 40% (4/10); Sto. Sunday, 50% (6/12); and Talugtug 50% (10/20). The seropositivity rate of N. caninum and the presence of T. evansi in Nueva Ecija may contribute to the cases of abortions in the province and further studies should be employed to confirm the association of these organisms to abortion cases on water buffaloes

Sensitive detection of Mycobacterium bovis in spiked milk using polymerase chain reaction assay

Bovine tuberculosis is a chronic zoonotic disease that affects both animal and human health and imposes serious public health concerns in the world. Intake of non-pasteurized milk is considered the most probable vehicle for the transmission of pathogenic bacteria. In this study, the detection of Mycobacterium bovis BCG in spiked milk using a polymerase chain reaction was performed. The performance of two DNA extraction methods, CTAB/phenol:chloroform:isoamyl alcohol and EXTRAGENMB were also evaluated. In addition, Mycobacterial concentration was tried to determine using the Standard/ Viable Plate Count Method and Spectrophotometric (Turbidimetric) Method. PCR successfully detected M. bovis BCG in spiked milk, detecting approximately up to two bacilli per reaction. The two DNA extraction methods were effective in the isolation of amplifiable DNA, having the advantage of EXTRAGENMB in terms of (1) shorter duration of DNA extraction, (2) less sample manipulation, and (3) ease of execution of the procedure. Quantitative determination of the Mycobacterial population however failed to quantify the bacterial concentration per dilution, suggesting that CFU concentration should be considered an approximation. It is expected that this method can be used for the detection of M. bovis in raw milk samples

Genotyping and Molecular Detection of Polymorphism in FUT1 Gene of Swine

Alpha (1, 2)-fucosyltransferase (FUT1), as a candidate gene in controlling the expression of E. coli F18 receptor has been identified to determine whether an animal is resistant or susceptible to infections. This study was conducted to determine the genotypes of 150 blood samples of the three swine breeds and 20 representatives were randomly selected for sequencing. Polymerase Chain Reaction - Restriction Fragment Length Polymorphism results revealed that among the 150 blood samples, seven of the Duroc x Pietrain samples carried AG genotype which was previously reported susceptible to ETEC infection. Two AA genotypes that are presumptively resistant and 50 were presumptively susceptible as these samples carried either AG or GG genotypes from the Landrace variety. Of the Large White samples, two samples carried AA genotype and 89 have either AG or GG genotype. The twenty samples sent for DNA sequencing registered 100% homologies to Sus scrofa FUT1 gene. Twelve of the sequenced samples exhibited a shift from G to A in the 117th nucleotide and one sample had a C to T shift in the 39th nucleotide. The mutation was found in all of the Duroc X Pietrain samples as well as those samples with AA genotype. Heterozygous forms of Landrace and Large White also exhibited this mutation

Evaluation of LAMP for detection and/or screening of Leptospira spp. infection among domestic animals in the Philippines

Objective: This study assessed the applicability of loop-mediated isothermal amplification (LAMP) for the detection of leptospirosis among domesticated animals and sewage rats. Specifically, it evaluated the ability of LAMP to amplify Leptospira spp. targeting the 16s rRNA gene in boiled urine samples. Materials and methods: A total of 140 samples from different domestic animals were tested for the presence of the antigen. A nested-polymerase chain reaction (nPCR) protocol was used to compare and determine the sensitivity of LAMP in detecting Leptospira spp. The LAMP was also evaluated by comparing its amplification result using agarose gel electrophoresis and color change using dye. Results: Positivity rate of Leptospira spp. antigen was 29.0% (40/140) for LAMP and 9.3% (13/140) for nPCR. Also, LAMP results for gel electrophoresis and dye color change varied in some samples that may be due to the interpretation of the result in dye color change. Conclusion: Overall, LAMP is a rapid, sensitive, and cost-effective diagnostic method compared with nPCR. Also, LAMP has a potential application as pen-side screening, surveillance, and clinical diagnostic kits of infectious diseases without requiring advance equipment and skilled personnel. [J Adv Vet Anim Res 2018; 5(4.000): 459-465