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Genetic diversity of Asian water buffalo (Bubalus bubalis): mitochondrial DNA D-loop and cytochrome b sequence variation

Lau CH, Drinkwater RD, Yusoff K, Tan SG, Hetzel DJ, Barker JS

Anim. Genet., 1998 Aug;29(4):253-64.
PMID: 9745663

Swamp and river buffalo mitochondrial DNA (mtDNA) was sequenced for 303 bp of the cytochrome b gene for 54 animals from 14 populations, and for 158 bp of the D-loop region for 80 animals from 11 populations. Only one cytochrome b haplotype was found in river buffalo. Of the four haplotypes identified in swamp buffalo, one found in all populations is apparently ancestral both to the other swamp haplotypes and to the river haplotype. The phylogenetic relationships among the 33 D-loop haplotypes, with a cluster of 11 found in swamp buffalo only, also support the evolution of domesticated swamp and river buffalo from an ancestral swamp-like animal, most likely represented today by the wild Asian buffalo (Bubalus arnee). The time of divergence of the swamp and river types, estimated from the D-loop data, is 28,000 to 87,000 years ago. We hypothesise that the species originated in mainland south-east Asia, and that it spread north to China and west to the Indian subcontinent, where the rive type evolved and was domesticated. Following domestication in China, the domesticated swamp buffalo spread through two separate routes, through Taiwan and the Philippines to the eastern islands of Borneo and Sulawesi, and south through mainland south-east Asia and then to the western islands of Indonesia.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Molecular evidence and hematological alterations associated with the occurrence of coronavirus in domestic dogs in Pakistan

Sulehria MU, Ahmad SS, Ijaz M, Mushtaq MH, Khan AY, Ghaffar A

Trop Biomed, 2020 Dec 01;37(4):963-972.
PMID: 33612749 DOI: 10.47665/tb.37.4.963

Canine Enteric Coronavirus (CCoV) is one of the major enteric pathogen affecting dogs. This study aims to investigate the molecular prevalence, phylogenetic analysis, associated risk factors, and haemato-biochemical alterations in Canine Coronavirus in dogs in district Lahore, Pakistan. 450 fecal samples were collected from symptomatic dogs originating from various pet-clinics and kennels during 2018-2019. Samples were initially analyzed by sandwich lateral flow immunochromatographic assay and then further processed by RT-PCR (reverse transcriptase polymerase chain reaction) targeting the M gene followed by sequencing. RT-PCR based positive (n=20) and negative (n=20) dogs were samples for their blood for the haemato-biochemical analysis. A questionnaire was used to collect data from pet owners, in order to analyze the data for risk factors analysis by chi square test on SPSS. The prevalence of CCoV was 35.1%, and 23.8 % through Sandwich lateral flow immunochromatographic and RT-PCR respectively. Various risk factors like breed, age, sex, vomiting, diarrhea, sample source, body size, cohabitation with other animals, living environment, food, deworming history, contact with other animals or birds feces, and season were significantly associated with CCoV. The CCoV identified in Pakistan were 98% similar with the isolates from China (KT 192675, 1), South Korea (HM 130573, 1), Brazil (GU 300134, 1), Colombia (MH 717721, 1), United Kingdom (JX 082356, 1) and Tunisia (KX156806). Haematobiochemical alterations in CCoV affected dogs revealed anaemia, leucopenia, lymphopenia, neutrophilia, and decreased packed cell volume, and a significant increase in alkaline phosphate and alanine transaminase. It is concluded that infection with canine coronavirus appears widespread among dog populations in district Lahore, Pakistan. This study is the first report regarding the molecular detection and sequence analysis of CCoV in Pakistan.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary
Fulltext Blood meal analysis of tabanid fly after it biting the rare Sumatran rhinoceros

Rovie-Ryan JJ, Zainuddin ZZ, Marni W, Ahmad AH, Ambu LN, Payne J

Asian Pac J Trop Biomed, 2013 Feb;3(2):95-9.
PMID: 23593586 DOI: 10.1016/S2221-1691(13)60031-3

To demonstrate a noninvasive large mammalian genetic sampling method using blood meal obtained from a tabanid fly.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Fulltext The effects of polymorphisms in 7 candidate genes on resistance to Salmonella Enteritidis in native chickens

Tohidi R, Idris IB, Malar Panandam J, Hair Bejo M

Poult Sci, 2013 Apr;92(4):900-9.
PMID: 23472012 DOI: 10.3382/ps.2012-02797

Salmonella enterica serovar Enteritidis infection is a common concern in poultry production for its negative effects on growth as well as food safety for humans. Identification of molecular markers that are linked to resistance to Salmonella Enteritidis may lead to appropriate solutions to control Salmonella infection in chickens. This study investigated the association of candidate genes with resistance to Salmonella Enteritidis in young chickens. Two native breeds of Malaysian chickens, namely, Village Chickens and Red Junglefowl, were evaluated for bacterial colonization after Salmonella Enteritidis inoculation. Seven candidate genes were selected on the basis of their physiological role in immune response, as determined by prior studies in other genetic lines: natural resistance-associated protein 1 (NRAMP1), transforming growth factor β3 (TGFβ3), transforming growth factor β4 (TGFβ4), inhibitor of apoptosis protein 1 (IAP1), caspase 1 (CASP1), lipopolysaccharide-induced tumor necrosis factor (TNF) α factor (LITAF), and TNF-related apoptosis-inducing ligand (TRAIL). Polymerase chain reaction-RFLP was used to identify polymorphisms in the candidate genes; all genes exhibited polymorphisms in at least one breed. The NRAMP1-SacI polymorphism correlated with the differences in Salmonella Enteritidis load in the cecum (P = 0.002) and spleen (P = 0.01) of Village Chickens. Polymorphisms in the restriction sites of TGFβ3-BsrI, TGFβ4-MboII, and TRAIL-StyI were associated with Salmonella Enteritidis burden in the cecum, spleen, and liver of Village Chickens and Red Junglefowl (P < 0.05). These results indicate that the NRAMP1, TGFβ3, TGFβ4, and TRAIL genes are potential candidates for use in selection programs for increasing genetic resistance against Salmonella Enteritidis in native Malaysian chickens.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Detection of Hepatocystis sp. in southeast Asian flying foxes (Pteropodidae) using microscopic and molecular methods

Olival KJ, Stiner EO, Perkins SL

J Parasitol, 2007 Dec;93(6):1538-40.
PMID: 18314711 DOI: 10.1645/GE-1208.1

Three species of flying fox (Pteropus hypomelanus, P. vampyrus, and P. lylei) from Malaysia and Vietnam were screened for apicomplexan parasites by thin blood smears and polymerase chain reaction. Only 1 of 16 bats sampled from 3 localities in southeast Asia was found to be infected (P. hypomelanus from Pulau Pangkor, Malaysia). We observed micro- and macrogametocytes, with morphology consistent with Hepatocystis sp. parasites, using light microscopy. Phylogenetic analysis of the cytochrome b gene showed that the parasite from P. hypomelanus groups with 2 published sequences from Hepatocystis spp., including one from Cynopterus brachyotis, another fruit bat in the Pteropodidae.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Intestinal spirochaetes (Brachyspira spp.) colonizing flocks of layer and breeder chickens in Malaysia

Amin MM, Phillips ND, La T, Robertson ID, Hampson DJ

Avian Pathol, 2014;43(6):501-5.
PMID: 25246135 DOI: 10.1080/03079457.2014.966056

Avian intestinal spirochaetosis causes problems including delayed onset of lay and wet litter in adult chickens, and results from colonization of the caecae/rectum with pathogenic intestinal spirochaetes (genus Brachyspira). Because avian intestinal spirochaetosis has not previously been studied in South East Asia, this investigation was undertaken in Malaysia. Faecal samples were collected from 25 farms and a questionnaire was administered. Brachyspira species were detected by polymerase chain reaction in 198 of 500 (39%) faecal samples from 20 (80%) farms, including 16 (94%) layer and four (50%) breeder farms. Pathogenic Brachyspira pilosicoli was identified in five (29%) layer and two (25%) breeder farms whilst pathogenic Brachyspira intermedia was detected in nine (53%) layer and one (12.5%) of the breeder farms. Twelve (80%) layer farms had egg production problems and 11 (92%) were positive for Brachyspira: three (25%) for B. pilosicoli and six (50%) for B. intermedia. Of three breeder farms with egg production problems, one was colonized with B. pilosicoli. Three of ten layer farms with wet litter were positive for B. pilosicoli and six for B. intermedia. Of four breeder farms with wet litter, one was colonized with B. pilosicoli and one with B. intermedia. No significant associations were found between colonization and reduced egg production or wet litter, perhaps because so many flocks were colonized. A significant association (P = 0.041) occurred between a high prevalence of colonization and faecal staining of eggs. There were significant positive associations between open-sided housing (P = 0.006), and flocks aged >40 weeks (P < 0.001) and colonization by pathogenic species.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Fulltext Detection of sarcocystosis in goats in Malaysia by light microscopy, histology, and PCR

Kutty MK, Latif B, Muslim A, Hussaini J, Daher AM, Heo CC, et al.

Trop Anim Health Prod, 2015 Apr;47(4):751-6.
PMID: 25740651 DOI: 10.1007/s11250-015-0789-4

A number of methods have been used for the detection of the presence of microsarcocysts in animals, but little information exists on the value between the various methods. This study therefore examined for Sarcocystis spp. using three different methods in 105 samples of skeletal muscle collected from goats slaughtered in an abattoir in Selangor, Malaysia from January to February 2014. Three methods were used, direct light microscopy of squashed fresh muscle tissues; histological examination of fixed, sectioned, and hematoxylin and eosin (H&E)-stained samples of muscle; and molecular identification by polymerase chain reaction (PCR). Of the 105 tissue samples, 55 (52.38 %) were positive by light microscopy (LM), 46 (43.8 %) by histology, and 95 (90.48 %) by PCR. Only 29 (27.6 %) and 5 (4.76 %) samples were positive and negative, respectively, by all three methods. The cysts were elongated to a spindle shape with a mean size of 393.30 × 81.6 μm and containing banana-shaped bradyzoites of size 12.32 × 2.08 μm. The wall of the cyst was radially striated with a thickness of 2.83 μm. Samples were tested for the presence of Sarcocystis-specific 18S rRNA and were identified as Sarcocystis capracanis. Of the three methods used, the PCR test appears to be the most useful method for the diagnosis of sarcocystosis especially for species identification.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Prevalence and antibiotic resistance of Salmonella Enteritidis and Salmonella Typhimurium in raw chicken meat at retail markets in Malaysia

Thung TY, Mahyudin NA, Basri DF, Wan Mohamed Radzi CW, Nakaguchi Y, Nishibuchi M, et al.

Poult Sci, 2016 Aug 01;95(8):1888-93.
PMID: 27118863 DOI: 10.3382/ps/pew144

Salmonellosis is one of the major food-borne diseases in many countries. This study was carried out to determine the occurrence of Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in raw chicken meat from wet markets and hypermarkets in Selangor, as well as to determine the antibiotic susceptibility profile of S. Enteritidis and S. Typhimurium. The most probable number (MPN) in combination with multiplex polymerase chain reaction (mPCR) method was used to quantify the Salmonella spp., S. Enteritidis, and S. Typhimurium in the samples. The occurrence of Salmonella spp., S. Enteritidis, and S. Typhimurium in 120 chicken meat samples were 20.80%, 6.70%, and 2.50%, respectively with estimated quantity varying from <3 to 15 MPN/g. The antibiogram testing revealed differential multi-drug resistance among S. Enteritidis and S. Typhimurium isolates. All the isolates were resistance to erythromycin, penicillin, and vancomycin whereas sensitivity was recorded for Amoxicillin/Clavulanic acid, Gentamicin, Tetracycline, and Trimethoprim. Our findings demonstrated that the retail chicken meat could be a source of multiple antimicrobial-resistance Salmonella and may constitute a public health concern in Malaysia.

Matched MeSH terms: Multiplex Polymerase Chain Reaction/veterinary
New Genus of Blood Fluke (Digenea: Schistosomatoidea) from Malaysian Freshwater Turtles (Geoemydidae) and its Phylogenetic Position Within Schistosomatoidea

Roberts JR, Platt TR, Orélis-Ribeiro R, Bullard SA

J Parasitol, 2016 08;102(4):451-62.
PMID: 27042972 DOI: 10.1645/15-893

: Baracktrema obamai n. gen., n. sp. infects the lung of geoemydid turtles (black marsh turtle, Siebenrockiella crassicollis [type host] and southeast Asian box turtle, Cuora amboinensis ) in the Malaysian states of Perak, Perlis, and Selangor. Baracktrema and Unicaecum Stunkard, 1925 are the only accepted turtle blood fluke genera having the combination of a single cecum, single testis, oviducal seminal receptacle, and uterine pouch. Baracktrema differs from Unicaecum by having a thread-like body approximately 30-50× longer than wide and post-cecal terminal genitalia. Unicaecum has a body approximately 8-12× longer than wide and terminal genitalia that are anterior to the distal end of the cecum. The new genus further differs from all other accepted turtle blood fluke genera by having a cecum that is highly convoluted for its entire length, a spindle-shaped ovary between the cirrus sac and testis, a uterine pouch that loops around the primary vitelline collecting duct, a Laurer's canal, and a dorsal common genital pore. Phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) revealed, with high nodal support and as predicted by morphology, that Baracktrema and Unicaecum share a recent common ancestor and form a clade sister to the freshwater turtle blood flukes of Spirorchis, paraphyletic Spirhapalum, and Vasotrema and that, collectively, these flukes were sister to all other tetrapod blood flukes (Hapalorhynchus + Griphobilharzia plus the marine turtle blood flukes and schistosomes). Pending a forthcoming emended morphological diagnosis of the family, the clade including Spirorchis spp., paraphyletic Spirhapalum, Vasotrema, Baracktrema, and Unicaecum is a likely placeholder for "Spirorchiidae Stunkard, 1921 " (type genus Spirorchis MacCallum, 1918 ; type species Spirorchis innominatus Ward, 1921 ). The present study comprises the 17th blood fluke known to infect geoemydid turtles and the first proposal of a new genus of turtle blood fluke in 21 yr.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
First report of Leishmania tropica in domestic and wild animal hosts in hyperendemic areas of human cutaneous leishmaniasis in western Yemen: a neglected tropical disease needing One Health approach

Al-Ashwal MA, Al-Adhroey AH, Atroosh WM, Al-Subbary AA, Albhri AA, Azlan UW, et al.

Parasitol Res, 2024 Jun 27;123(6):256.
PMID: 38935203 DOI: 10.1007/s00436-024-08273-3

Cutaneous leishmaniasis (CL), a neglected tropical disease, is a major public health concern in Yemen, with Leishmania tropica identified as the main causative agent. This study aims to investigate the occurrence and distribution of Leishmania parasites in domestic and wild animals in CL endemic areas in the western highlands of Yemen. A cross-sectional study was conducted in the Utmah District of western Yemen. Blood and skin scraping specimens were collected from 122 domestic and wild animals and tested for the Leishmania DNA using internal transcribed spacer 1 (ITS1) nested polymerase chain reaction. Phylogenetic analyses were performed on 20 L. tropica sequences obtained from animals in this study and 34 sequences from human isolates (collected concurrently from the same study area) retrieved from the GenBank. Overall, L. tropica was detected in 16.4% (20/122) of the examined animals, including 11 goats, two dogs, two bulls, one cow, one donkey, one rabbit, one rat and one bat. None of the examined cats and sheep was positive. The animal sequences were segregated into four different L. tropica haplotypes, with the majority of the animal (15/20) and human (32/34) sequences composed of one dominant haplotype/genotype. These findings represent the first confirmed evidence of natural L. tropica infections in different kinds of domestic and wild animals in western Yemen, suggesting these animals potentially have a role in the transmission of CL in Yemen. Therefore, a One Health approach is required for the effective prevention and control of this devastating disease among endemic populations.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Tracing the origins of genotype VIIh Newcastle disease in southern Africa

Abolnik C, Mubamba C, Wandrag DBR, Horner R, Gummow B, Dautu G, et al.

Transbound Emerg Dis, 2018 Apr;65(2):e393-e403.
PMID: 29178267 DOI: 10.1111/tbed.12771

It is widely accepted that Newcastle disease is endemic in most African countries, but little attention has been afforded to establishing the sources and frequency of the introductions of exotic strains. Newcastle disease outbreaks have a high cost in Africa, particularly on rural livelihoods. Genotype VIIh emerged in South-East Asia and has since caused serious outbreaks in poultry in Malaysia, Indonesia, southern China, Vietnam and Cambodia. Genotype VIIh reached the African continent in 2011, with the first outbreaks reported in Mozambique. Here, we used a combination of phylogenetic evidence, molecular dating and epidemiological reports to trace the origins and spread of subgenotype VIIh Newcastle disease in southern Africa. We determined that the infection spread northwards through Mozambique, and then into the poultry of the north-eastern provinces of Zimbabwe. From Mozambique, it also reached neighbouring Malawi and Zambia. In Zimbabwe, the disease spread southward towards South Africa and Botswana, causing outbreaks in backyard chickens in early-to-mid 2013. In August 2013, the disease entered South Africa's large commercial industry, and the entire country was infected within a year, likely through fomites and the movements of cull chickens. Illegal poultry trading or infected waste from ships and not wild migratory birds was the likely source of the introduction to Mozambique in 2011.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary
Fulltext Pathological changes and bacteriological assessments in the urinary tract of pregnant goats experimentally infected with Brucella melitensis

Mazlina M, Khairani-Bejo S, Hazilawati H, Tiagarahan T, Shaqinah NN, Zamri-Saad M

BMC Vet Res, 2018 Jun 25;14(1):203.
PMID: 29940976 DOI: 10.1186/s12917-018-1533-x

BACKGROUND: This study was conducted to investigate the pathological changes and distribution of B. melitensis in the urinary tract of pregnant goats following acute experimental infection. Six Jamnapari crossbred does in their third trimester of pregnancy were randomly assigned into two groups; Group 1 was uninfected control and Group 2 was inoculated conjunctival with 0.1 mL of the inoculums containing 109 cfu/mL of live B. melitensis. All does were sacrificed 30 days post-inoculation before the kidney, ureter, urinary bladder, urethra and vaginal swab were collected for isolation of B. melitensis. The same tissue samples were fixed in 10% neutral buffered formalin for hematoxylin and eosin, and immunoperoxidase staining.
RESULTS: None of the goats showed clinical signs or gross lesions. The most consistent histopathology finding was the infiltration of mononuclear cells, chiefly the macrophages with few lymphocytes and occasionally neutrophils in all organs along the urinary tract of the infected goats of Group 2. Other histopathology findings included mild necrosis of the epithelial cells of the renal tubules, congestion and occasional haemorrhages in the various tissues. Kidneys showed the most severe lesions. Immunoperoxidase staining revealed the presence of B. melitensis within the infiltrating macrophages and the epithelium of renal tubules, ureter, urethra and urinary bladder. Most extensive distribution was observed in the urinary bladder. Brucella melitensis was successfully isolated at low concentration (3.4 × 103 cfu/g) in the various organs of the urinary tract and at high concentration (2.4 × 108 cfu/mL) in the vaginal swabs of all infected goats. Although B. melitensis was successfully isolated from the various organs of the urinary tract, it was not isolated from the urine samples that were collected from the urinary bladder at necropsy.
CONCLUSION: This study demonstrates the presence of low concentrations of B. melitensis in the organs of urinary tract of pregnant does, resulting in mild histopathology lesions. However, B. melitensis was not isolated from the urine that was collected from the urinary bladder.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Fulltext Hexon and fiber gene changes in an attenuated fowl adenovirus isolate from Malaysia in embryonated chicken eggs and its infectivity in chickens

Sohaimi NM, Bejo MH, Omar AR, Ideris A, Isa NM

J Vet Sci, 2018 Nov 30;19(6):759-770.
PMID: 30173491 DOI: 10.4142/jvs.2018.19.6.759

Fowl adenovirus (FAdV) is distributed worldwide and causes economic losses in the poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate from Malaysia in specific pathogen-free chicken embryonated eggs (SPF CEE) and its infectivity in commercial broiler chickens. SPF CEE were inoculated with 0.1 mL FAdV inoculum via the chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20), with a virus titer of 108.7TCID50/mL (TCID50, 50% tissue culture infective dose), was inoculated (0.5 mL) into one-day-old commercial broiler chicks either via oral or intraperitoneal routes. The study demonstrated that 100% embryonic mortality was recorded from E2 to E20 with a delayed pattern at E17 onwards. The lesions were confined to the liver and CAM. Substitutions of amino acids in the L1 loop of hexon at positions 49 and 66, and in the knob of fiber at positions 318 and 322 were recorded in the E20 isolate. The isolate belongs to serotype 8b and is non-pathogenic to broiler chickens, but it is able to induce a FAdV antibody titer. It appears that molecular changes in the L1 loop of hexon and the knob of fiber are markers for FAdV infectivity.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Genomic analysis of some Japanese isolates of Getah virus

Wekesa SN, Inoshima Y, Murakami K, Sentsui H

Vet Microbiol, 2001 Nov 08;83(2):137-46.
PMID: 11557154

Using the reverse transcription-polymerase chain reaction (RT-PCR) and direct sequencing, capsid protein and non-structural protein 1 (nsP1) regions of Sagiyama virus and eight Getah virus strains were analysed. The viruses were isolated from Malaysia and various areas of Japan over a period of 30 years. Based on the available published sequence data, oligonucleotide primers were designed for RT-PCR and the sequences were determined. Our findings showed that though there were differences in the nucleotide sequences in the nsP1 region, there was 100% amino acid homology. On the other hand, in the capsid region, the nucleotide differences caused a major difference in the amino acid sequence. Therefore, the difference in the capsid region is one of the useful markers in the genetic classification between Sagiyama virus and strains of Getah virus, and might be responsible for the serological difference in complement fixation test. The genomic differences among the Getah virus strains are due to time factor rather than geographical distribution.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary
Molecular characterization of Salmonella weltevreden isolated from poultry: evidence of conjugal transfer of plasmid and antibiotic resistance

Radu S, Mutalib SA, Rusul G, Hassan Z, Yeang LK

Microbios, 2001;104(407):39-47.
PMID: 11229656

Ten strains of Salmonella weltevreden isolated from poultry sources were examined and found to contain plasmid DNA ranging in size from 1.8 to 68.5 MD. All isolates were susceptible to carbenicillin, cephalothin, ceftriazone, gentamicin, kanamycin and nalidixic acid, but resistance to bacitracin (100%), penicillin G (100%), rifampicin (100%), sulphamethoxazole (100%), cefuroxime (80%) and tetracycline (60%) was recorded. The 55 MD plasmid of strain SW5 determined resistance to penicillin G and tetracycline, which was transmissible to the E. coli K12 recipient at a frequency of 3.52 x 10(-5) transconjugants per input donor cell. The results of arbitrarily primed polymerase chain reaction (AP-PCR), using two 10-mer oligonucleotides and PCR-ribotyping to differentiate between the ten strains of S. weltevreden were compared. The strains were separated into ten different genome types by AP-PCR but were indistinguishable by PCR-ribotyping. These results suggest that poultry may constitute a reservoir for disseminating antibiotic resistance and that AP-PCR may be a valuable tool for epidemiological studies.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Molecular characterization of a Toxocara variant from cats in Kuala Lumpur, Malaysia

Zhu XQ, Jacobs DE, Chilton NB, Sani RA, Cheng NA, Gasser RB

Parasitology, 1998 Aug;117 ( Pt 2):155-64.
PMID: 9778638

The ascaridoid nematode of cats from Kuala Lumpur, Malaysia, previously identified morphologically as Toxocara canis, was characterized using a molecular approach. The nuclear ribosomal DNA (rDNA) region spanning the first internal transcribed spacer (ITS-1), the 5.8S gene and the second internal transcribed spacer (ITS-2) was amplified and sequenced. The sequences for the parasite from Malaysian cats were compared with those for T. canis and T. cati. The sequence data showed that this taxon was genetically more similar to T. cati than to T. canis in the ITS-1, 5.8S and ITS-2. Differences in the ITS-1 and ITS-2 sequences between the taxa (9.4-26.1%) were markedly higher than variation between samples within T. canis and T. cati (0-2.9%). The sequence data demonstrate that the parasite from Malaysian cats is neither T. canis nor T. cati and indicate that it is a distinct species. Based on these data, PCR-linked restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP) methods were employed for the unequivocal differentiation of the Toxocara variant from T. canis and T. cati. These methods should provide valuable tools for studying the life-cycle, transmission pattern(s) and zoonotic potential of this parasite.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Encephalitis and retarded growth of chicks caused by Sitiawan virus, a new isolate belonging to the genus Flavivirus

Kono Y, Tsukamoto K, Abd Hamid M, Darus A, Lian TC, Sam LS, et al.

Am J Trop Med Hyg, 2001 5 19;63(1-2):94-101.
PMID: 11358004

A new virus named Sitiawan virus (SV) was isolated from sick broiler chicks in chicken embryos. The virus replicated well with cytopathogenic effect (CPE) in the chicken B-lymphocyte cell line LSCC-BK3. The virus was an enveloped RNA virus of approximately 41 nm in size with hemagglutinating activity (HA) to goose erythrocytes. It was cross-reactive with Japanese encephalitis virus (JEV), a member of flaviviruses by HA inhibition tests but not by cross-virus neutralization tests. The cDNA fragment of NS5 gene was amplified with primers corresponding to NS5 gene of flaviviruses. The nucleotide sequences were 92% homologous to Tembusu virus, a member of the mosquito-borne virus cluster of the genus Flavivirus. In cross-neutralization tests with Tembusu virus, antiserum to SV did not neutralize Tembusu virus, and antiserum to Tembusu virus neutralized more weakly to SV than against homologous virus. These results indicate that SV is a new virus which can be differentiated serologically from Tembusu virus but is otherwise similar with respect to nucleotide sequence. The virus causes encephalitis, growth retardation, and increased blood glucose levels in inoculated chicks.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary
Iridovirus disease in two ornamental tropical freshwater fishes: African lampeye and dwarf gourami

Sudthongkong C, Miyata M, Miyazaki T

Dis Aquat Organ, 2002 Apr 5;48(3):163-73.
PMID: 12033703

Many species of ornamental freshwater fishes are imported into Japan from all over the world. We found African lampeye Aplocheilichthys normani and dwarf gourami Colisa lalia suffering from an iridovirus infection just after being imported by tropical fish wholesalers from Singapore. African lampeye were cultured on the Indonesian Island of Sumatra and dwarf gourami were cultured in Malaysia before export. Diseased fishes displayed distinct histopathological signs of iridovirus infection: systemic appearance of inclusion body-bearing cells, and necrosis of splenocytes and hematopoietic cells. Electron microscopy revealed viral particles (African lampeye:180 to 200 nm in edge to edge diameter; dwarf gourami: 140 to 150 nm in diameter) in an inclusion body within the cytoplasm of inclusion body-bearing cells as well as in the cytoplasm of necrotized cells. Experimental infection with an iridovirus isolate from African lampeye (ALIV) revealed pathogenicity of ALIV to African lampeye and pearl gourami Trichogaster leeri. Polymerase chain reaction (PCR) products from ALIV and an iridovirus isolate from dwarf gourami (DGIV) using iridovirus-specific primers were indistinguishable. The nucleotide sequence of PCR products derived from ALIV (696 base pairs) and DGIV (701 base pairs) had 95.3% identity. These results indicate that ALIV and DGIV have a single origin.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
MOLECULAR INVESTIGATION AND PHYLOGENETIC ANALYSIS OF ANAPLASMOSIS IN DOGS

Ghauri HN, Ijaz M, Ahmed A, Muhammad Naveed MUA, Nawab Y, Javed MU, et al.

J Parasitol, 2021 03 01;107(2):295-303.
PMID: 33844841 DOI: 10.1645/20-50

Anaplasmosis is a widespread vector-borne disease affecting dogs, and Anaplasma platys is the major etiological agent of the disease. The study examines anaplasmosis molecular prevalence, related risk factors, and alteration of hematological variables in Anaplasma-affected dogs. A total of 150 blood samples were collected from dogs in the district of Lahore, Pakistan. The samples were screened with PCR targeting the 16S rRNA gene of Anaplasma. Sequencing of samples that were found positive after performing PCR was conducted. A questionnaire was developed to collect epidemiological data on subject dogs, and the information was analyzed with a logistic regression model using SPSS. The current study revealed an 11.34% (17/150) prevalence of anaplasmosis in dogs based on PCR detection. Tick infestation, previous tick history, house hygiene, and tick control status were major risk factors linked with disease occurrence. Red blood cell count, packed cell volume, hemoglobin, and platelet count were decreased significantly (P < 0.05) in Anaplasma-infected dogs. Phylogenetically, the 2 isolates of the current study clustered together, and that cluster was very similar to A. platys isolates from India, Malaysia, and Thailand.

Matched MeSH terms: Polymerase Chain Reaction/veterinary
Fulltext Co-administration of avian influenza virus H5 plasmid DNA with chicken IL-15 and IL-18 enhanced chickens immune responses

Lim KL, Jazayeri SD, Yeap SK, Alitheen NB, Bejo MH, Ideris A, et al.

BMC Vet Res, 2012;8:132.
PMID: 22866758 DOI: 10.1186/1746-6148-8-132

DNA vaccines offer several advantages over conventional vaccines in the development of effective vaccines against avian influenza virus (AIV). However, one of the limitations of the DNA vaccine in poultry is that it induces poor immune responses. In this study, chicken interleukin (IL) -15 and IL-18 were used as genetic adjuvants to improve the immune responses induced from the H5 DNA vaccination in chickens. The immunogenicity of the recombinant plasmid DNA was analyzed based on the antibody production, T cell responses and cytokine production, following inoculation in 1-day-old (Trial 1) and 14-day-old (Trial 2) specific-pathogen-free chickens. Hence, the purpose of the present study was to explore the role of chicken IL-15 and IL-18 as adjuvants following the vaccination of chickens with the H5 DNA vaccine.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary

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