Identification of Rodent Species That Infest Poultry Houses in Mafikeng, North West Province, South Africa

Rodents cause serious adverse effects on farm production due to destruction of food, contamination of feed, and circulation of diseases. The extent of damage or the diseases spread will depend on the type of rodents that invade the farm. This study was conducted in order to find out the species of rodents that infest poultry farms around Mafikeng, North West Province of South Africa. The study was part of a broader project that was investigating Salmonella vectors in the poultry farms around the province. The study trapped 154 rodents from selected farms and used the Cytochrome oxidase subunit 1 (COI) and the Cytochrome b (Cyt-b) barcoding genes for species identification. Two rodent pest species, namely, Rattus rattus (the black rat) and Rattus tanezumi (the Asian Rat/Asian House Rat) were identified. A total of 99 (64.3%) were identified as Rattus rattus and 55 (35.7%) were Rattus tanezumi. Between the two target genes, Cyt-b gene was only able to identify 40 (25.97%) of the total samples while COI was more efficient and amplified all the samples and thus was a better target gene for this kind of identification. The two rat species identified are known vectors of serious diseases; thus their presence should be regarded as an indication of high risk for diseases. Despite having been detected in the country before, finding R. tanezumi as the second largest rat species in the area was unexpected since this species is known to be indigenous to Asia

Use of reverse transcriptase loop-mediated isothermal amplification assay for field detection of Newcastle disease virus using less invasive samples

A novel nucleic acid amplification method, loop-mediated isothermal amplification, was developed and recently demonstrated detection of Newcastle disease virus (NDV) in tissue samples. But slaughter of poultry for test samples is often faced with resentment by low-income farmers. This study was undertaken to determine the test properties of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) in detection of NDV in clinical cases using cloacal and oropharyngeal swabs. Samples included 46 tracheal tissues, 94 cloacal and 107 oro-pharyngeal swabs from on-station and 30 spleens, 74 cloacal and 74 oro-pharyngeal swabs from the field. Analysis was done using specific RT-LAMP targeting the fusion (F) protein. While the method detected NDV from swab samples, no RNA of other poultry disease viruses was amplified, indicating analytical specificity of 100%. RT-LAMP took ≤36 minutes in 83% (n=329) of positive reactions with all samples amplified in <60 minutes. Results were easily observed with a naked eye. Cloacal and oro-pharyngeal swabs could be a convenient and cheaper alternative in diagnosis of NDV infection by RT-LAMP in resource poor countries. [Vet. World 2012; 5(4.000): 206-212

First report of gastrointestinal nematodes and coccidia parasites from free-range chickens in Mafeteng district, Lesotho

Free-range chickens are an integral part of poultry production in many developing countries. In the Mountain Kingdom of Lesotho, the majority of the population own free-range chickens, which serve a variety of purposes including being a source of meat, eggs and use for cultural rituals amongst others. However, there is lack of scientific studies on occurrence of parasitic infections on free-range chickens in Lesotho. The aim of this study was to document common gastrointestinal parasites infecting free-range chickens in four villages of Mafeteng District in Lesotho. A total number of 462 pooled faecal samples were collected from various households in HaKubutu (n = 114), HaMatjeka (n = 120), HaMpalipali (n = 120) and Thabang Villages (n = 108) which were subjected to microscopic examination using McMaster technique. The prevalence of gastrointestinal parasite infection was as follows: Eimeria tenella (12.8%), Ascaridia galli (10.4%) and Heterakis gallinarum (5%). The prevalence for H. gallinarum and Ascaridia galli were comparatively higher during the hot-wet season (7.1% and 2.8% respectively) than the cold-dry season (3.2% and 1.9% respectively) and varied significantly (P < 0.05). For E. tenella, the oocysts per gram were slightly higher in the cold-dry season than the hot-wet season. Polymerase chain reaction only amplified DNA from six (29%) adult A. galli worms of which two amplicons were successfully sequenced. The obtained cytochrome C oxidase subunit 1 partial gene sequences displayed 98–100% identity with South African A. galli isolates. This is the first scientific study on prevalence and molecular characterization of nematodes and coccidia species infecting free-range village chickens in Lesotho. The findings can be used to review management of gastrointestinal nematodes and protozoal parasites of free-range chickens in Lesotho

Molecular detection and genetic characterisation of pathogenic Theileria, Anaplasma and Ehrlichia species among apparently healthy sheep in central and western Kenya

Tick-borne diseases (TBDs) caused by Theileria, Babesia, Anaplasma and Ehrlichia species are common in tropical and subtropical regions. In this study, we investigated the presence and genetic diversity of Theileria spp., Anaplasma ovis, B. ovis, E. ruminantium and Anaplasma spp. in sheep from the Machakos and Homa Bay counties of Kenya. In order to improve the diagnosis and control of ovine TBDs, a total of 76 blood samples from apparently healthy sheep were screened using a polymerase chain reaction (PCR). The assays were conducted using primers based on Theileria spp. 18S rRNA, Anaplasma ovis Major surface protein-4 (AoMSP4), B. ovis 18S rRNA, E. ruminantium pCS20 and Anaplasma spp. 16S rRNA. The overall infection rates for Theileria spp., A. ovis, E. ruminantium and Anaplasma spp. were 39/76 (51.3%), 26/76 (34.2%), 6/76 (7.9%) and 31/76 (40.8%), respectively. The overall co-infection was 47/76 (61.8%). All Theileria spp. positive samples were confirmed to be of Theileria ovis on sequencing. A phylogenetic analysis of the 18S rRNA gene sequences of T. ovis revealed that all isolates of this study clustered with T. ovis sequences extracted from the GenBank suggesting this gene is highly conserved. E. ruminantium pCS20 sequences were in the same clade on the phylogenetic tree. However, three AoMSP4 sequences from this study appeared in the same clade, while one sequence formed a separate branch revealing genetic divergence. The 16S rRNA sequencing revealed uncharacterised Anaplasma spp. and A. ovis. The phylogenetic analyses of the uncharacterised Anaplasma spp. revealed that the two sequences from this study appear in an independent clade from other sequences extracted from the GenBank. This study provides important information regarding the occurrence of tick-borne pathogens and their degree of genetic diversity among sheep in Kenya, which is useful for the diagnosis and control of TBDs

Theileria in ruminants

Theileria are important hemoprotozoan parasites of domestic and wild ruminants, transmitted by ixodid ticks leading to diseases which range from mild in apparent reactions to highly fatal diseases. Bovine-infecting T. parva and T. annulata and ovine-infecting T. lestoquardi are of major global economic importance, but other Theileria spp. are also briefly mentioned. Classification of Theileria has been a subject of great controversy without consensus on whether many taxons are different species, synonyms, or subspecies of the same parasite. However, with the development of new molecular tools, many of the outstanding difficulties could be resolved. Theileria have complex life cycles both in the vertebrate host and the tick vector, many of which are not clearly understood. One unique feature of some Theileria is their ability to transform infected host cells into a reversible cancer-like proliferation conferring to them an ability to proliferate without apoptosis. The transformation is not permanent because it can be reversed by treatment with anti-theilerial drugs. Understanding this mechanism could give insights into treatment of cancer. Control of the diseases caused by Theileria has largely relied on chemical drugs either to treat infected hosts or prevent infection by controlling the tick vectors. But resistance to chemicals by the parasites or vectors has led to the development of more sustainable control methods such as live vaccination against the three most pathogenic Theileria spp. of ruminants. Efforts are also under way to develop subunit vaccines against these parasites