Active infection and morphometric study of Trypanosoma evansi among horses in Peninsula Malaysia.

Apart from occasional reports of clinical disease affecting horses, there is no information about Trypanosoma evansi in horses in Peninsula Malaysia. Thus, a cross-sectional study was conducted in eight states in Peninsula Malaysia to determine the active presence of T. evansi in horses. A total of 527 blood samples were obtained and examined by haematocrit centrifugation technique (HCT), Giemsa-stained thin blood smear (GSS), morphometric measurements, polymerase chain reaction (PCR) and cloning of PCR products. The results showed an overall parasitological prevalence of 0.57% (3/527, CI: 1.6-0.19%) with both HCT and GSS. Morphometric study revealed the mean total length of the trypanosomes including the free flagellum was 27.94 ± 2.63 µm. PCR successfully amplified a trypanosome specific 257 bp in 1.14% of samples (6/527, CI: 2.4-0.52%) and was confirmed by nucleotide sequences. The mean packed cell volume (PCV) for the positive cases detected by HCT was lower (23% ± 7.00) compared to the positive cases detected by PCR alone in the state of Terengganu (35% ± 4.73). In conclusion, this study showed T. evansi infection occurred in low frequency in horses in Peninsula Malaysia, and anaemia coincided with parasitaemic animals. PCR is considered as a sensitive diagnostic tool when parasitaemia is undetectable. The slight lengthier mean of parasite and anaemia may indicate a virulent strain of T. evansi circulating throughout the country. Thus, it's highly recommended to shed light on host-parasite relationship for better epidemiological understanding.

Evaluation of enhanced virgin coconut oil and senduduk (Melastoma malabathricum) as anthelmintics against caprine strongyle nematodes.

A study was conducted to evaluate the anthelmintic properties of enhanced virgin coconut oil (EVCO) and senduduk (Melastoma malabathricum) plant against strongyle nematodes in goats. Two preparations of 10% EVCO dissolved in 90% virgin coconut oil and 10% EVCO dissolved in 90% palm oil, were given orally to two groups of mixed breeds goats. The efficacy test indicated that EVCO was insufficiently active as an anthelmintic. Four concentrations of senduduk solution (1.25, 2.5, 5.0 and 10 mg ml(-1)) were compared with a control and albendazole in an in vitro test for larvicidal effect. There was no significant larval mortality using senduduk solution. An in vivo test of senduduk was conducted by comparing three groups of goats, namely control, levamisole and treatment groups that were given a daily oral dose of senduduk crude extract with 1g kg(-1) from Day 0 to Day 12 and 2 g kg(-1) from Day 13 to Day 30. This efficacy test with senduduk also gave negative results. The findings obtained indicated that EVCO and senduduk were ineffective as anthelmintics against caprinestrongyle nematodes at the concentrations used.

Seroprevalence and risk factors of Trypanosoma evansi infection in horses in Peninsular Malaysia.

A cross-sectional study was designed to assess the seroprevalence and risk factors associated with Trypanosoma evansi infection among horses, using a total of 527 blood samples obtained from eight states in Peninsular Malaysia. A structured questionnaire was used to collect data on risk factors associated with T. evansi seroprevalence. The overall seroprevalence detected by card agglutination test for T. evansi (CATT/T. evansi) was 13.90% (73/527, CI: 11.2-17.1%). Female and exogenous horses showed a higher risk in association with the disease seroprevalence compared to other groups. The majority of the horse owners were not familiar with surra (85.30%). However, most of them were very cautious with the health of their animals. In conclusion, this study showed that T. evansi occurred in low frequency among horses in Peninsular Malaysia, and the good management system adopted by horse owners was probably responsible for the low T. evansi occurrence.

Low prevalence of Dirofilaria immitis in dogs in Johor Bahru, Malaysia as a reflection of vector availability?

This study was conducted to investigate the low prevalence of Dirofilaria immitis in dogs in Johor Bahru as reported by veterinary practitioners, using wet blood mount, Knott's Concentration Test and two heartworm antigen test kits (IDEXX Canine SNAP® 4Dx and RapiGEN®). This study also compared the two test kits used and determined the microfilaria species. Blood were collected from 100 owned dogs and 50 stray dogs in Johor Bahru via cephalic venipuncture. A thick blood smear was done and examined for samples that were positive for microfilaria species identification. The overall prevalence of D. immitis in dogs in Johor Bahru was 1.33% (2/150) and the microfilaria identified was D. immitis. The prevalence of heartworm in owned and stray dogs in this study was 1% and 2% respectively. With only one false negative result from RapiGEN® test kit, comparing the sensitivity between the two test kits could not be achieved. The low prevalence of D. immitis found in this study confirmed anecdotal evidence that prevalence of dirofilariasis is indeed low in Johor Bahru. Additionally, we speculate that dirofilariasis in dogs might be considered as an indicator of vector availability.

Low prevalence of Dirofilaria immitis in dogs in Johor Bahru, Malaysia as a reflection of vector availability?

Abstract. This study was conducted to investigate the low prevalence of Dirofilaria immitis in dogs in Johor Bahru as reported by veterinary practitioners, using wet blood mount, Knott’s Concentration Test and two heartworm antigen test kits (IDEXX Canine SNAP® 4Dx and RapiGEN®). This study also compared the two test kits used and determined the microfilaria species. Blood were collected from 100 owned dogs and 50 stray dogs in Johor Bahru via cephalic venipuncture. A thick blood smear was done and examined for samples that were positive for microfilaria species identification. The overall prevalence of D. immitis in dogs in Johor Bahru was 1.33% (2/150) and the microfilaria identified was D. immitis. The prevalence of heartworm in owned and stray dogs in this study was 1% and 2% respectively. With only one false negative result from RapiGEN® test kit, comparing the sensitivity between the two test kits could not be achieved. The low prevalence of D. immitis found in this study confirmed anecdotal evidence that prevalence of dirofilariasis is indeed low in Johor Bahru. Additionally, we speculate that dirofilariasis in dogs might be considered as an indicator of vector availability.

Lungworm of cattle in Malaysia.

Bovine lungworm Dictyocaulus viviparus is highly endemic in temperate regions. However, the occurrence of the lungworm has not been reported in any South East Asian country. The main aim of the present study was to detect the presence of lungworm in cattle in peninsular Malaysia and to examine the morphology of the parasite. A cross-sectional study was carried out in which 602 animals from four large scale government cattle farms and one dairy smallholder farm were sampled. In addition, 283 lungs from 11 abattoirs around the country were examined. Faecal samples were examined using the Baermann technique while post-mortem examination was performed on the lungs. Approximately 5% of faecal samples and 1% of lungs were positive for lungworm. Based on the morphology of adult lungworm, eggs and first stage larvae, Malaysian bovine lungworms were D. viviparus.

Screening of microfilaricidal effects of plant extracts against Dirofilaria immitis.

Canine dirofilariasis is a common tropical parasitic disease of companion animals, caused by infestation of Dirofilaria immitis filarids within the pulmonary arteries and extending into the right heart. Increased reports of adverse reactions elicited by current microfilaricidal agents against D. immitis such as neurological disorders, circulatory collapse and potential resistance against these agents, warrant the search for new agents in forms of plant extracts. The use of plant extracts in therapeutic medicine is commonly met with scepticism by the veterinary community, thus the lack of focus on its medical potential. This study evaluated the presence of microfilaricidal activities of the aqueous extracts of Zingiber officinale, Andrographis paniculata and Tinospora crispa Miers on D. immitisin vitro at different concentrations; 10mg/ml, 1mg/ml, 100 microg/ml, 10 microg/ml and 1 microg/ml within 24h, by evaluation of relative microfilarial motility as a measure of microfilaricidal activity. All extracts showed microfilaricidal activity with Z. officinale exhibiting the strongest activity overall, followed by A. paniculata and T. crispa Miers. It is speculated that the microfilaricidal mechanism exhibited by these extracts is via spastic paralysis based upon direct observation of the microfilarial motility.

Endo-parasite fauna of rodents caught in five wet markets in Kuala Lumpur and its potential zoonotic implications.

Rodents were collected from five wet markets (Chow Kit, Dato Keramat, Setapak, Jinjang and Kepong) in Kuala Lumpur, Federal Territory between March to April 2006. Ninety seven rats were trapped using wire traps measuring 29 x 22 x 50 cm baited with fruits, coconuts, dried fish or sweet potatoes. A total of 17 different species of parasites were identified from three species of rats out of which 11 (65%) were identified to be zoonotic. The helminths identified from the urban rats were nematodes- Capillaria hepatica, Gongylonema neoplasticum, Heterakis spumosa, Heterakis sp., Masterphorus muris, Nippostrongylus brasiliensis, Physolaptera sp., Pterogodermatis sp., Rictularia tani and Syphacia muris; cestodes- Hymenolepis nana, Hymenolepis diminuta, Hymenolepis sabnema, Hymenolepis sp., Raillietina sp. and Taenia taeniaeformis, and acanthocephalan- Moniliformis moniliformis. The following parasites are of potential medical importance: C. hepatica, G. neoplasticum, R. tani, S. muris, H. diminuta, H. nana, Raillietina sp. and T. taeniaeformis.

Ectoparasite fauna of rodents and shrews from four habitats in Kuala Lumpur and the states of Selangor and Negeri Sembilan, Malaysia and its public health significance.

A total of 204 rodents comprising 14 host species from four different habitats were examined. Nine rodent species were trapped from the forest and another five species were trapped from the coastal, rice field and urban habitats. Rattus rattus diardii (67%) was the predominant rodent species examined. Fifty six (47.3%) rodents and shrews were found to be infested with at least one of the 20 species of ectoparasite recovered. Mites belonging to the family Trombiculidae were the predominant ectoparasite species recovered. Ticks belonging to the family Ixodidae were recovered mainly from the forest dwelling rodents. Polyplax spinulosa and Hoplopleura pacifica were the common lice species found infesting the urban rodents. Xenopsylla cheopis was the only flea species recovered. The following ecto-parasites have been incriminated as important vectors or as mechanical carriers for the transmission of zoonotic diseases: Ixodes granulatus, Dermacentor sp. Haemaphysalis sp., Amblyomma sp. Ascoschoengastia indica, Leptotrombidium deliense, Ornithonyssus bacoti, Laelaps nuttalli, H. pacifica, P. spinulosa and Xenopsylla cheopis. Urban and forest rodents were significantly higher in ecto-parasitic infestation, compared to rats from the other two habitats. However, there was no significant statistical association between male and female rodents infested with ectoparasites.

First report of bovine lungworm disease in South-East Asia.

This paper presents investigation of lungworm disease outbreaks that is based on retrospective examination of cases recorded between 1994 and 2000 on a government beef cattle breeding centre in the state of Pahang, peninsular Malaysia. The breed of cattle on the centre was Nelore and the mean population over a 7-year period (from 1994 to 2000) was 1612. All animals were allowed to graze on pasture and mixed grazing was practiced on the farm. The routine de-worming programme was performed using levamisole and ivermectin from 1994 to 1998 and abamectin in 1999 and 2000 on 1 to 3-month-old calves and an annual dose given to the adult cattle. Nelore was introduced into the farm in 1991, three years before the first outbreak from Brazil where Dictyocaulus viviparus infection had been reported. No lungworm infection had been observed in the farm prior to the animal introduction. Within the 7-year period, 36 fatalities occurred and the annual mortality rate due to lungworm infection was 0.31%. The highest rate was recorded in 1997. Among the total 36 deaths, about 75% of deaths occurred in calves aged between 6 months and 12 months, 67% were males and 33% were female cattle. The highest number of deaths (19%) occurred in the month of November. In conclusion, D. viviparus infection may have been introduced into a tropical climate along with consignments of cattle from lungworm endemic areas resulting in fatal disease outbreaks for a few years following the animal's initial introduction.

PCR tools for the verification of the specific identity of ascaridoid nematodes from dogs and cats.

Based on the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of Toxocara canis, Toxocara cati, Toxocara malaysiensis and Toxascaris leonina, specific forward primers were designed in the ITS-1 or ITS-2 for each of the four ascaridoid species of dogs and cats. These primers were used individually together with a conserved primer in the large subunit of rDNA to amplify partial ITS-1 and/or ITS-2 of rDNA from 107 DNA samples from ascaridoids from dogs and cats in China, Australia, Malaysia, England and the Netherlands. This approach allowed their specific identification, with no amplicons being amplified from heterogeneous DNA samples, and sequencing confirmed the identity of the sequences amplified. The minimum amounts of DNA detectable using the PCR assays were 0.13-0.54ng. These PCR assays should provide useful tools for the diagnosis and molecular epidemiological investigations of toxocariasis in humans and animals.

A comparative longitudinal study of bovine trypanosomiasis in tsetse-free and tsetse-infested zones of the Amhara Region, northwest Ethiopia.

A study was conducted to determine the incidence of trypanosome infections in cattle in tsetse-free and tsetse-infested zones of the Amhara Region of northwest Ethiopia. A total of six sentinel herds were established and the cattle observed during a period of 8 consecutive months. The prevalence of seropositive cattle was high in both the tsetse-free and tsetse-infested zones. The average monthly incidence of trypanosome infection, determined using molecular diagnostic tools, was 20.9% and 25.7% in the tsetse-free and the tsetse-infested zones, respectively. In the tsetse-free, Trypanosoma vivax was responsible for 90.9% of the cattle trypanosome infections. In the tsetse-infested zone, Trypanosoma congolense and T. vivax contributed almost equally to the trypanosome infections in cattle. Trypanosome infection, regardless of species, resulted in anaemia as evidenced by a significant decrease in the packed cell volume of the infected animal. The outcome of this longitudinal study suggests that control of trypanosomiasis in the Amhara Region cannot be achieved by tsetse control alone. Supplemental measures to include drug therapy and biting fly control are discussed.

Isolation of Neospora caninum from a calf in Malaysia.

In order to attempt isolate the protozoan parasite Neospora caninum, an N. caninum seropositive pregnant Sahiwal Friesian cross heifer from a large-scale dairy farm in Malaysia was kept for observation until parturition at the Veterinary Research Institute, Ipoh. The heifer gave birth to a female calf that was weak, underweight and unable to rise. Precolostral serum from the calf had an N. caninum indirect fluorescent antibody test titre of 1:3200. It died 12 h after birth and necropsy was performed. Brain homogenate from the calf was inoculated into 10 BALB/c mice that were kept for 3 months after which brain tissue from the mice was inoculated onto 24 h fresh monolayer Vero cell lines. The cell cultures were examined daily until growth of intracellular protozoa was observed. DNA of the organisms from the cell cultures was analyzed by PCR and DNA sequencing. DNA fragments of the expected size were amplified from the isolate using N. caninum-specific primers, and sequence analysis of ITS1 clearly identified the isolate as N. caninum. This is the first successful isolation of N. caninum from a bovine in Malaysia, and the isolate is designated Nc-MalB1.

Seasonal prevalence of bovine trypanosomosis in a tsetse-infested zone and a tsetse-free zone of the Amhara Region, north-west Ethiopia.

During a period of four consecutive years, trypanosomosis surveys were conducted in a tsetse-infested and tsetse-free area of the Amhara Region of north-west Ethiopia. In each study area randomly selected communal cattle were sampled and their blood was investigated using parasitological diagnostic methods. At the same time the population of biting flies was sampled. The monthly average prevalence of trypanosome infections in cattle did not differ significantly between study areas. In both study areas, the prevalence of trypanosome infections was highest during the long rainy season. Trypanosome infections were mainly due to Trypanosoma vivax and they significantly reduced the average packed cell volume and the body condition of the animals. The monthly prevalence of infection was correlated with the density of biting flies, such as Tabanidae and Stomoxys spp., in the preceding month suggesting an important role of mechanical transmission in the epidemiology of trypanosomosis in both areas.

Redescription of Serpinema octorugatum (Baylis, 1933) nematoda: Camallanidae from the Malayan box turtle Cuora amboinensis Daudin Chelonia: Bataguridae.

We redescribe the camallanid nematode Serpinema octorugatum (Baylis, 1933) from the box turtle Cuora amboinensis (Daudin) collected in Malaysia. In this redescription, we amend the original description by noting that there are only four cephalic papillae and that there are five pairs of post-anal papillae, and propose that the name of this species be corrected from S. octorugatus to S. octorugatum. Additionally, we removed the tissues overlying the buccal capsule and have used SEM studies to show that the peribuccal shields extend laterally from the buccal capsule, forming a surface possibly used in muscle attachment. Furthermore, we show that the supposedly non-cuticularised cylinder connecting the buccal capsule to the oesophagus in the Camallanidae is part of the buccal capsule and is, therefore, likely to be cuticularised. We also examine morphological measurements of taxonomic interest for correlations with total body length and find that many characters traditionally used for inter- and intra-specific comparisons are correlated with total body length in adult female worms. This suggests that comparisons between samples of adult female worms that do not account for the potential effect of total body length may be misleading. However, we show that some features of taxonomic interest are not correlated with total body length.

Toxocara malaysiensis n. sp. (Nematoda: Ascaridoidea) from the domestic cat (Felis catus Linnaeus, 1758).

Toxocara malaysiensis n. sp. from the small intestine of the domestic cat (Felis catus L.) in Malaysia is described and illustrated. This ascaridoid nematode was previously assumed to be Toxocara canis, which it superficially resembles, or designated Toxocara sp. cf. canis. The new species differs from T. canis in the shape of the cervical alae in cross section, spicule length, and the lip structure. It is also distinct from other species assigned to Toxocara.

Epidemiology of Trypanosoma evansi infection in crossbred dairy cattle in Malaysia.

An investigation into the epidemiology of Trypansoma evansi infection in crossbred dairy cattle was conducted for a period of 12 months on a dairy cattle farm in Penninsular Malaysia. The prevalence of parasitaemia was highest in lactating animals (13.4%), followed by those in the dry herd (8.8%), late pregnant animals (8.1%), early pregnant animals (4.7%), calves (0.3%) and heifers (0.2%). The prevalence of antigenaemia was highest in the lactating animals (54.7%), followed by that in dry animals (53.7%), heifers (51.1%), late pregnant animals (47.7%), early pregnant animals (46.5%) and calves (24.2%).

Molecular characterization of a Toxocara variant from cats in Kuala Lumpur, Malaysia.

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.

Dietary protein influences on regulation of Haemonchus contortus populations in Dorsimal lambs.

The influence of dietary protein supplementation upon resistance to haemonchosis was examined in Dorsimal (Polled Dorset x Malin) lambs offered two levels of protein. Lambs were offered either a complete basal ruminant diet (15% crude protein (CP)) or the same diet supplemented with fish meal as a source of rumen bypass protein (19% CP). Lambs from each dietary treatment group were given either a 7-week trickle infection with Haemonchus contortus infective larvae (L3) or remained uninfected. All lambs were drenched with anthelmintic at week 8 post-infection (PI), challenged with a single dose of 5000 H. contortus L3 one week later, and killed 14 days post-challenge (PC). Lambs on the supplemented diet that were trickle infected showed a significant reduction in egg output. Supplementation and previous infection did not affect either growth rate, worm burden, worm development or haematological parameters. There was a trend for enhanced growth among supplemented non-infected lambs in comparison to lambs which received the basal ration.