Evidence of novel Treponema phylotypes implicated in contagious ovine digital dermatitis and association of treponemes with major lameness causing foot pathogens.

In this study 269 swabs collected from 254 ovine foot lesions and 15 apparently healthy ovine feet were screened by PCR for the presence of major lameness causing foot pathogens viz. Treponema species, D. nodosus, F. necrophorum and T. pyogenes with the presumption that ovine foot lesion positive for Treponema species alone or in association with other three pathogens were categorized as contagious ovine digital dermatitis (CODD). While samples positive for D. nodosus alone or its combination with F. necrophorum and T. pyogenes were considered as footrot (FR) and samples in which F. necrophorum or T. pyogenes was found either alone or in combination were considered as interdigital dermatitis (ID). The overall occurrence of Treponema sp. in ovine foot lesions was 48.0%, and ranged from 33 to 58%. In Treponema positive samples D. nodosus, F. necrophorum and T. pyogenes were present in 34 (27.4%), 66 (54.4%) and 84 (68.5%) in contrast to Treponema negative samples in which these were present in 15 (11.1%), 20 (14.12%) and 17 (12.6%) samples, respectively. The data signifies that Treponema sp. are significantly associated with these foot pathogens and their different combinations with Treponema sp. influence the severity of CODD lesion. The identification of Treponema phylotypes was done by sequencing the 16S rRNA gene fragment of ten representative samples. Out of ten sequences, four (Trep-2, Trep-4, Trep-7 and Trep-10) were identical to Treponema sp. phylotype 1 (PT1) that belongs to phylogroup T. refringens-like, one sequence (Trep-1) was genetically close (90% sequence homology) to Treponema brennaborense while five sequences (Trep-3, Trep-5, Trep-6, Trep-8 and Trep-9) matched with uncultured bacterium clones of treponemes forming separate monophyletic group in phylogenetic tree and could represent new digital dermatitis phylogroup presently containing five ovine specific phylotypes. This is the first report on the presence of Treponema phylotypes other than three digital dermatitis (DD) Treponema phylogroups viz. T. phagedenis-like, T. medium/T. vincentii-like, and T. pedis-like that are frequently detected in CODD lesions. Metagenomic analysis of two representative samples revealed the abundance of genus Treponema in CODD lesion while this genus was absent in swab collected from clinically healthy foot suggesting that it might play primary role in producing CODD. These findings may further aid in understanding the etiopathogenesis of CODD and could help to develop appropriate treatment and mitigation strategies to combat the disease.

Factors influencing seedling emergence of three global invaders in greenhouses representing major eco-regions of the world.

Successful germination and seedling emergence in new environments are crucial first steps in the life history of global plant invaders and thus play a key role in processes of range expansion. We examined the germination and seedling emergence success of three global plant invaders - Lupinus polyphyllus, Senecio inaequidens and Verbascum thapsus - in greenhouses and climate chambers under climate regimes corresponding to seven eco-regions. Seed materials were collected from one non-native population for L. polyphyllus and S. inaequidens, and from 12 populations for V. thapsus (six natives and six non-natives). Experimental climates had significant effects on species responses. No species germinated in the dry (humidity ≤ 50%) and cool (≤ 5 °C) experimental climates. But all species germinated and emerged in two moderately cool (12-19 °C) and in three warm (24-27 °C) experimental climates. In general, V. thapsus showed higher fitness than S. inaequidens and L. polyphyllus. The climate of the seed source region influenced responses of native and non-native populations of V. thapsus. Non-native populations of V. thapsus, originating from the warmer seed source, showed higher performance in warm experimental climates and lower performance in moderately cool experimental climates compared to native populations. Responses of V. thapsus populations were also related to precipitation of the seed source region in moderately dry experimental climates. The warm, semi-arid and humid experimental climates are suitable for the crucial first steps of invasion success for L. polyphyllus, S. inaequidens and V. thapsus. The species adaptation to its source region modified the responses of our studied plants under different experimental climates representing major eco-regions of the world.

Sindbis virus replicase-based DNA vaccine construct encoding FMDV-specific multivalent epitope gene: studies on its immune responses in guinea pigs.

Foot-and-mouth disease (FMD) is still a perennial global menace affecting livestock health and production. It is imperative to figure out new ways to curb this disease. In this study, a sindbis virus replicase-based DNA vaccine, pSinCMV-Vac-MEG990, encoding a multivalent epitope gene (representing tandemly linked VP1 C-terminal halves of three foot-and-mouth disease virus (FMDV) serotypes) was constructed. In vitro transfection studies in BHK-21 cells revealed that the construct was able to express FMDV-specific antigen but does not overproduce the antigen. Immunization of guinea pigs with the construct at dose rate of 10, 5, 2 and 1 µg per animal through intramuscular route showed significant neutralizing antibody induction at all doses against all serotype tested as compared to non-immunized controls. On viral challenge of guinea pigs 4 week post-immunization with 1000 GPID(50) of FMDV serotype A, it was observed that the immunization not only delayed the appearance and reduced the severity of FMD lesions significantly (P < 0.05) but also provided complete protection in several guinea pigs. In fact, two of six and one of six guinea pigs were completely protected in 10 and 5 µg immunized groups, respectively. These results suggest that the development of the replicase-based DNA vaccine may provide a promising approach as an alternative vaccine strategy for controlling FMD.

Intestinal helminths in a population of children from the Kashmir valley, India.

In any geographical area, surveys of the prevalence of intestinal helminths are necessary to suggest appropriate control measures. The aim of this study was to determine the prevalence of intestinal helminth infections in children of the Kashmir valley and to identify the risk factors. Stool samples were collected from 2256 children from rural as well as urban areas of the Kashmir valley. The samples were examined by simple smear and zinc sulphate concentration methods. Intensity of the infection was quantified by Stoll's egg-counting technique. Infection by at least one intestinal helminth was found in 71.18% of the sampled population. The prevalence of Ascaris lumbricoides was highest (68.30%), followed by Trichuris trichiura (27.92%), Enterobius vermicularis (12.67%) and Taenia saginata (4.60%). Light (57.1%) to moderate (42.8%) intensity of infection was observed for A. lumbricoides, while the majority of the infected children (92.3%) harboured a light intensity of infection for T. trichiura. The age group, rural or urban residence, type of water source, boiled or unboiled water, type of defecation site, level of personal hygiene and maternal education were associated with helminth infection. Adequate control measures are urgently needed to combat the high prevalence of intestinal helminths and risk factors in the children of Kashmir valley.