Detection of Glasser's Disease in clinical samples using Polymerase chain reaction.

Glasser’s Disease is one of the porcine common respiratory problems in out country. Glasser’s disease is caused by Haemophilus parasuis (HP). The HP has a wide range of antibiotics sensitivity and can be treated during early infection; therefore early diagnosis is very important. In the past, diagnosis was based on history, clinical signs and postmortem lesions, then confirm by bacteria isolation and identification. These traditional microbiology methods are time-consuming and labourious. Polymerase chain reaction (PCR) is one of the methods that provide rapid and accurate diagnosis. The procedure can be completed within 1 to 2 days and only require one trained personal to perform. Currently, there is limited publish articles on PCR test for the diagnosis of Glasser’s Disease in Malaysia. For the analysis 2 pairs of primers set for HP were selected; HPS1-forward (5’AGT AGG AAG GGT GGT GT 3’) and HPSI-reverse (5’ CGT TTC GTC ACC CTC TGT GT 3’) and HPS2- forward (5’ TAG AAA AAA TCT TTA ATT G 3’) and HPS2-reverse (5’ CAC CAT AGA AAC TTC TTT TC 3’). Lung tissues, pericardial swabs and thoracic swabs samples were collected form farms in Sepang, Selangor, Malaysia. The PCR test was carried after some modifications and optimization. Both HP primers able to detect positive clinical lung samples and can be further developed as PCR diagnosis tool in out country

Detection of mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome in clinical samples by polymerase chain reaction

The aim of this study was detect Swine Enzootic Pneumonia (SEP) and Porcine Reproductive and Respiratory Syndrome (PRRS) from clinical affected lung samples with PCR technique. In this technique three primers set used for M. hyopneumonieae detection were Cai (Forward: GAG CCT TCA AGC TIC ACC AAG A: Reverse: TGT GTT AGT GAC TTT TGC CAC C), Baumeister (Forward: TAG AAA TGA CTG GCA GAC AA: Reverse: GAG GCT TGA TTT TGG AGT C) and Caron (Forward: GAC CCG ATG AAA CCT ATT AAA ATA GAC: Reverse; GAA GCG AAA TTA AAT ATT TTT AAT TCG ATA CTG). For the detection of PRRS virus the primer sets used Oleksievicz (Forward: GAA CCT GCC CAI CAC G: Reverse: TAC CCC TAA TTG AAT AGG GGA) amd Suàrez )Forward: GGG AAT GGC CAG CCA GTC AAT CAA CTG T: Reverse: TGT AGA AGT CAC GCG AAT CAG GCG CAC T). Nine pigs aged betwee 6-10 weeks were collected from farms in Selangor, Malaysia. One healthy pig and 8 other pigs with clinical signs of respiratory distress problem were sacrificed and lung bronchoalveolar lavage samples were obtained. Healthy pigs were selected as negative control while samples were harvested from 4 pigs with suspected SEP and 4 with respiratory problems for M. hyoneumoniae and PRRS virus detection. Base on the result, the Caronand Caiprimer sets were able to detect SEP from the affected lungs. For PRRS virus, RNA was extracted using easy-BLUE™ Total RNA Extraction Kit and converted it to cDNA with Maxime RT PreMix Kit. The oleksiewicz primer set was ideally suited for the detection of PRRS virus

Effect of simultaneous injection of classical swine fever virus vaccine and Mycoplasma hyopneumoniae vaccine on immune response of swine

Objectives of this study were (1) to compare sero-conversion in pigs following simultaneous and separate vaccination against Classical Swine Fever (CSF) and Mycoplasma hyopneumoniae and (2) to determine safety of CSF and M. hyopneumoniae vaccines when given simultaneously. Twenty-four weaned pigs were divided into 3 groups of 8 heads. Groups were designated as non-simultaneous vaccinated group, simultaneous vaccinated group and negative control, respectively. Vaccines used in study were M.hyopneumoniae vaccine (SPRINTVAC®MH) and CSF vaccine (PESTIFFA®). IDEXX ELISA test kit (HerdChek M hyo) and LSIVET SUIS HC/PPC Blocking ELISA test kit were used to detect antibody titre on weekly basis. Sero-conversion rate of CSF antibody titre and M.hyo antibody titre were calculated. Result showed both simultaneous vaccination and non-simultaneous vaccination for CSF antibody titre reached 100% sero-conversion rate at 5 weeks post vaccination. Therefore, simultaneous vaccination was able to accomplish similar results as in non-simultaneous vaccination. Sero-conversion rate for CSF antibody titre in non-simultaneous group was slower before it reached 5 weeks post vaccination. 12.5% of animal from negative control group sero-converted at 5 weeks post vaccination due to false-positive result or field infections. M. hyopneumoniae antibody titre sero-conversion rate in both simultaneous vaccination and non-simultaneous vaccination reached 100% sero-conversion rate after 6 weeks post vaccination. Control group showed negative result for M. hyopneumoniae antibody titre throughout whole experiment. Vaccines used in trial did not cause any adverse effect after post vaccination when given simultaneously

Detection of the Japanese encephalitis virus in wild boars and comparison of blood profiles between wild boars and domestic pigs in Selangor, Malaysia

The Japanese encephalitis virus (JEV) RNA was detected in mosquito vectors in Selangor, Malaysia almost two decades ago. However, the JEV status in wild boars, a potential reservoir, has yet to be investigated. Blood profiles can be used for health monitoring however data on the wild boar blood profile is limited. This study was performed to detect the presence of JEV RNA in wild boars in Selangor, Malaysia and to determine the haematological and serum biochemical values of the wild boars. The blood profiles of domestic pigs were also determined for comparison. Thirty-five wild boar tonsils were collected for RNA extraction while 21 wild boar and 40 domestic pig blood were collected for analyses. The RNA extraction was performed using the QIAamp RNA Blood Mini Kit (Qiagen, USA) while reverse transcription and PCR amplification were performed using the i-JEV Detection Kit (iNtron Biotechnology, Korea). The blood analyses were performed using standard protocols in the Haematology and Clinical Biochemistry Laboratory, Faculty of Veterinary Medicine, Universiti Putra Malaysia. The JEV RNA (genotype I and/or III) was detected in one of 21 (4.8%) tonsils; gene sequencing can be done to plot a phylogenetic tree to identify the origin of virus. The positivity may be higher if a vigourously optimized multiplex RT-PCR was used in the detection JEV genotypes. The leucocyte, segmented neutrophil counts and serum globulin concentration of the wild boars were significantly (p<0.05) lower than in domestic pigs and the existing reference data. Serum AST and CK concentration of the wild boars were significantly higher than that of domestic pigs, which might be attributed the physically more active wild boars

A preliminary study of methicillin-resistant Staphylococcus aureus and antimicrobial resistance profile of bacteria in selected pig farms in Peninsular Malaysia

Staphylococcus aureus is one of the most common and devastating human and animal pathogens. In this study, conventional technique of isolation and identification was used to isolate Staphylococus aureus, Staphylococcus hyicus, Streptococcus spp., and E. coli. Detection and confirmation of MRSA was done using Staphytect® Kit and Oxacillin Resistance Screening Agar Base (ORSAB). Two farms from each pig industry rearing state namely Selangor, Perak, Johor and Penang were selected. Nasal and rectal swab samples were taken from 64 piglets and nasal swab samples were taken from 16 farm workers. Nineteen (13.2%) MRSA were isolated. Fourteen (21.9%) isolates were from pigs while 5 (31.3%) isolates originated from humans. These alarming findings from the present study indicated that MRSA is an emerging pathogen as well as a zoonotic potential in pigs and humans in Malaysia. This study also determined the antimicrobial sensitivity profile of some isolates towards commonly used antimicrobials in pig farms. Results showed that lincomycin is no longer effective for treatment in the farms while spectinomycin, florfenicol and enrofloxacin are starting to be less effective in controlling pathogens. Both colistin and ceftiofur are still effective as the bacteria tested are sensitive towards them. The findings from this study warrant that suitable measures must be undertaken to prevent the spread of MRSA as well as to control the rise of antibiotic resistant bacteria in the farms

Characterisation of porcine reproductive and respiratory syndrome virus strains in selected farms in Malaysia

Porcine reproductive and respiratory syndrome (PRRS) is a disease characterised by late-term reproductive failure in sows and gilts, and respiratory problems in piglets and growing pigs. The PRRS virus can be divided into two antigenically and genetically different strains: Type I (European) and Type II (North American). In this study, 120 sera were collected from 12 farms in 6 states in Malaysia for the seroprevalence study. Ten sera from apparently healthy sows/gilts, finishers and growers were collected from each farm and tested using IDEXX HerdChek ELISA for both strains. All the farms tested were seropositive with an overall seroprevalence of 89.2%. Tissue samples were collected and PRRSV isolated were genotyped using nested-PCR (with reported primer pairs targeting ORF7) that enabled the differentiation of Type I and Type II PRRSV by producing different sizes of PCR products. Out of 27 tissue samples collected from 11 farms, 12 were positive for PRRSV. All the PRRSV genomes from the 12 PRRSV-positive tissue homogenates were of Type II PRRSV, whereas no Type I PRRSV was detected. These data indicate that PRRS is endemic in the farms tested with a high possibility of subclinical infections

Melamin toxicity in pigs

Three farms in Penang, Malaysia, reported illness in grower-finisher pigs with discarded kidneys from abattoir. Affected animals were inappetant, anorexic, thin, dull, and depressed. The episode was closely related with the feed formulation changes, where soybean and fishmeal were replaced with yeast. No significant mycotoxin levels were detected in both the feed and yeasts. However, high levels of melamine were detected in yeast (30,064 ppm) and finished feed (750 to 1500 ppm), which exceeded the acceptance level set by WHO (<2.5 ppm). Nonetheless, prominent gross abnormalities were detected, except for the lesions in the kidneys during necropsy. The affected kidneys were enlarged and yellowish in colour, firm in consistency and the cortical surface was wrinkled and dimpled. Dilated cortical and medullary lesions with yellow gritty crystalline materials were also observed. A microscopic examination revealed the lesions of acute tubular nephrosis, interstitial nephritis with multiple cysts in the kidney cortex. After the removal of the yeast source in the feed, the illness and kidney lesions were in remission

Treatments of chicken feather waste

Feather waste is a potential renewable source to recover valuable products because it is being a rich source of keratin proteins and amino acids. It can be used to make feather meal, fertilizer and yarn sizing agent. Various treatments have been used to recover the protein from chicken feathers as the keratinous feathers cannot be easily degraded due to its tough structure. This paper reviews the existing treatment methods used to hydrolyze chicken feathers. The treatment methods for feather hydrolysis such as physical, chemical, biological and combined treatments as well as their advantages and challenges are highlighted. The effects of these treatments on feather hydrolysis are complex and vary in regards to the performance of feather hydrolysis and product yielded. Hence, it is important to choose an appropriate treatment method since the type of treatment applied affects the product yielded qualitatively and quantitatively. In addition, the economic assessment and environmental impact of the choice of treatment should be considered also

Investigative baseline reference on the status of pork pH, shear force, colour, drip and cooking loss in RYR1 mutation free, commercial 3-way crosses in Malaysia

This paper attempts to provide findings of an investigative study on the baseline status of the pork quality in Malaysia. With consumer preferences changing towards the selection of good quality meat for consumption, there is a need to establish an investigative reference for the operators in the industry to gauge the performance of their animals and pork quality. This is also important to increase the competitiveness among producers to continuously improve the pork quality available to consumers. In this study, 30 commercial three-way crossed female pigs were randomly selected from government accredited abattoirs from east and west Malaysia and longisimus dorsi were collected for the determination of pH, drip loss, cooking loss, shear force and colour. All animals were screened for the RYR1 gene and the results were then compiled with statistical analysis to obtain an investigative baseline pork quality data in Malaysia. The average pork quality obtained from this study falls within the category of Red, Soft and Exudative (RSE), with an average ultimate pH of 5.83, drip loss more than 5% and L* values at 45.94. We have proposed an investigative baseline meat quality data for Malaysian pork from the average commercial pork quality data obtained. The proposed investigative pork quality baseline data in Malaysian is comparable in terms of studies done in other established countries and/or with international standards and falls within the RSE category of acceptable quality. It provides an investigative benchmark for researchers and end-producers to judge the quality of pork in an objective manner, both for consumption and for export purpose. Moreover, continuous selection against the RYR1 gene has successfully removed the gene from the sample size above, but constant random monitoring is still advisable if farms aim to ensure the elimination of this gene from their herd

Detection of classical swine fever virus by a surface plasmon resonance assay

Test sera from vaccinated pigs at different time-frame were used to assess the sensitivity and specificity of an optimized Surface Plasmon Resonance (SPR)-chip-based detection method. Biomolecular interaction between Classical Swine Fever Virus (CSFV) and serum antibody were investigated in relative time using Biacore SPR system. An amount of 8860.93 RU of CSFV in 10mM sodium acetate of pH 5.0 was fixed on CM5 dextran matrix sensor chip. Serum from vaccinated animals was allowed to run over the whole CSFV in triplicate. The relative response from serum of 5th week and 7th week old swine towards the immobilized CSFV reacted variably. The 10 fold diluted serum response unit of 94.24 ± 11.34 RU indicated a limited immune response at week 5. However in week 7, the highest response in the serum at same dilution demonstrated a 2 fold increase at 189.33 ± 2.57 RU. Regeneration with glycine-HCl at pH 2.0 enabled successful baseline reversion after each analysis. The herein established, whole virus immobilized SPR-chip could serve as a prototype for a rapid and sensitive CSFV diagnosis assay