Comparing genetic diversity in agroforestry systems with natural forest: a case study of the important timber tree Vitex fischeri in central Kenya

It is possible that current tree domestication practices undertaken by farmers reduce the genetic base of tree resources on farms, raising concerns regarding the productivity, sustainability and conservation value of agroforestry ecosystems. Here, we assessed possible changes in genetic variation during domestication in the important and heavily utilised timber species, Vitex fischeri Gürke (syn. Vitex keniensis), by comparing geographically proximate forest and farm material in central Kenya. Employing RAPD analysis, a total of 104 polymorphic markers revealed by five arbitrary primers were scored in a total of 65 individuals, 32 from forest and 33 from farmland. Despite concerns of possible genetic erosion, forest and farm stands did not differ significantly in levels of genetic variation, with H values of 0.278 and 0.269, respectively. However, Mantel tests did reveal greater geographically related associative genetic structure among individuals in farm rather than forest material, with r _M values of 0.217 and 0.114, respectively. A more detailed analysis of structure suggested this could be due to local variation in origin of some on-farm trees. Implications of data for the genetic management of V. fischeri stands during farmer-led tree domestication activities are discussed. At present, there appears little reason to reject on-farm V. fischeri as a source of germplasm for future on-farm planting or for conservation purposes, although this situation may change and will require monitoring.     

A B-cell targeting virus disrupts potentially protective genomic methylation patterns in lymphoid tissue by increasing global 5-hydroxymethylcytosine levels

The mechanisms by which viruses modulate the immune system include changes in host genomic methylation. 5-hydroxymethylcytosine (5hmC) is the catalytic product of the Tet (Ten-11 translocation) family of enzymes and may serve as an intermediate of DNA demethylation. Recent reports suggest that 5hmC may confer consequences on cellular events including the pathogenesis of disease; in order to explore this possibility further we investigated both 5-methylcytosine (5mC) and 5hmC levels in healthy and diseased chicken bursas of Fabricius. We discovered that embryonic B-cells have high 5mC content while 5hmC decreases during bursa development. We propose that a high 5mC level protects from the mutagenic activity of the B-cell antibody diversifying enzyme activation induced deaminase (AID). In support of this view, AID mRNA increases significantly within the developing bursa from embryonic to post hatch stages while mRNAs that encode Tet family members 1 and 2 reduce over the same period. Moreover, our data revealed that infectious bursal disease virus (IBDV) disrupts this genomic methylation pattern causing a global increase in 5hmC levels in a mechanism that may involve increased Tet 1 and 2 mRNAs. To our knowledge this is the first time that a viral infection has been observed to cause global increases in genomic 5hmC within infected host tissues, underlining a mechanism that may involve the induction of B-cell genomic instability and cell death to facilitate viral egress.     

Linkage relationships among loci controlling morphological traits in cowpea (Vigna unguiculata (L.) Walp.)

Summary Linkage among loci controlling various morphological traits in cowpea were determined using F₂ progenies. Data were collected on individual plants of four crosses segregating for several loci. Recombination estimates between the following pairs of loci were as follows: Sw (swollen vs normal stem base)-Fbc (cream vs green flower buds) (41±4.8%), Pu^s(purple vs green stems)-Cbr (cocoa-brown vs straw-yellow dry pods) (31±5.7%), Pu^p(purple vs green immature pods)-Cbr (30±5.7%), Pu^s-Pu^p (4±1.5%), Ndt (non-determinate vs determinate)-Pd (peduncle colour) (26±2.8%), Ndt-Hg (semi-erect vs erect plant type) (26±2.8%), P^t(purple vs green pod tips)-Bk (greyish-black vs straw-yellow dry pods) (19±2.4%) and Hg-Bpd (normal vs branching peduncle (24±9.5%). Four linkage groups (LG) were identified in these studies. The proposed LG I contained loci Sw and Fbc; LG II loci Pu^s, Pu^p, and Cbr; LG III loci Pd, Ndt, Gh, and Bpd; and LG IV loci P^tand Bk.     

Genetics of resistance to Aphis craccivora in cowpea

Summary Inheritance of aphid resistance and allelic relationships among sources of resistance was studied in the parents, F₁, F₂, F₃, and backcross populations of cowpea crosses. Each 4-day old seedling was infested with five fourthinstar aphids. Seedling reaction was recorded 14–16 days after infestation when the susceptible check was killed. The segregation data from eight crosses between resistant and susceptible cowpea cultivars indicated that aphid resistance was inherited as a monogenic dominant trait. Segregation data from crosses among eight resistant cultivars indicated that one or two loci and modifier(s) were involved in the expression of resistance to aphids. It was suggested that further studies on allelism among sources of resistance needed to be conducted in order to resolve this.     

A study of the justification for intensive tick control in Kenyan rangelands

Sixty male weaner cattle at the National Range Research Station, Kiboko, Kenya were placed in four groups of 15 animals under four different tick control regimens for 16 months. The treatment groups were: spraying with acaricide weekly, spraying every three weeks, spraying whenever group mean tick infestations reached more than 200 per animal (once only in the course of the study) and a control unsprayed group. The cattle were weighed monthly. There were no significant differences between the liveweight gains of the groups during a period of severe drought and during the following period of compensatory weight gain. The untreated group gained more weight than the other groups. Cattle died in all groups, but without significant differences between the groups; the long drought and associated malnutrition were the prime cause of death. Rhipicephalus pravus and R pulchellus were the dominant tick species with fewer Amblyomma gemma, Hyalomma truncatum and small numbers of Boophilus decoloratus and R evertsi. These ticks transmit anaplasmosis, babesiosis and cowdriosis in the study area. Total tick counts reached 750 on individual animals but group means rarely reached more than 200 because of high resistance to ticks in some animals. Host resistance ranking within groups was virtually constant throughout the experiment. The study showed that intensive tick control is not required in the semi-arid areas of Africa.     

Risk factors associated with occurrence of nematodes in free range pigs in Busia District,Kenya

Nematode infections are a serious constraint to pig production, especially where free range pig keeping is practiced. This study investigated the epidemiology of nematodes in free range pigs in Busia District, Kenya. Three hundred and six pigs from 135 farms were sampled for faeces that were analysed for nematode eggs per gram (EPG) of faeces using the McMaster technique. The nematode eggs were also identified to genus and species based on morphology. A questionnaire on risk factors was also administered to the pig owners. The overall prevalence and mean nematode EPG were 84.2% and 2,355, respectively. The nematode eggs were identified as those belonging to Oesophagostomum spp. (75%), Strongyloides ransomi (37%), Ascaris suum (18%), Metastrongylus spp. (11%), Trichuris suis (7%) and Physocephalus sexalatus (3%). The prevalence of nematodes was positively correlated (p < 0.05) with the amount of rainfall in the division of the pigs' origin (all nematodes except S. ransomi). The prevalence of nematodes was also associated with the age of the pigs. A lower burden of nematodes was associated (p < 0.05) with a history of deworming (A. suum) and the provision of night housing (S. ransomi and Metastrongylus spp.). In conclusion, this study has provided information on nematode infections and the associated risk factors for free range pigs in Busia District, which can be used when implementing integrated control measures.     

Bovine NK cells acquire cytotoxic activity and produce IFN-gamma after stimulation by Mycobacterium bovis BCG- or Babesia bovis-exposed splenic dendritic cells

Early interactions of innate immune cell populations, such as dendritic cells (DC) and natural killer (NK) cells, can affect the ability of the acquired immune response to control infection of intracellular microorganisms. In this study, we investigated the activation of bovine NK cells by CD13(+) splenic DC stimulated with either Mycobacterium bovis BCG or Babesia bovis merozoites. Splenic DC were used either immediately after selection (cytokine(-)) or after exposure to GM-CSF, IL-4 and Flt3L for 72 h (cytokine(+)). Phenotypic analyses showed up-regulation of MHCII, CD80 and CD86 on cytokine(+) DC when compared to cytokine(-) DC. Purified NK cells (CD335(+)CD3(-)CD2(+/-)CD8alpha(+/-)) were co-cultured with microbial-exposed cytokine(-) DC or cytokine(+) DC in either transwell or cell-to-cell format and NK cell IFN-gamma production and cytotoxicity were assessed. NK cell IFN-gamma production was dependent on cell-to-cell contact. Microbial-stimulated cytokine(+) DC induced significantly more IFN-gamma production from NK cells than cytokine(-) cells. In contrast, cytotoxicity and perforin up-regulation were more pronounced in NK cells cultured with cytokine(-) DC than cytokine(+) DC. Therefore, activation of bovine NK cells by microbial-stimulated CD13(+) splenic DC is influenced by the maturation state of the DC suggesting different roles for the splenic DC during disease-induced maturation.     

Evaluation of the recognition of Theileria parva vaccine candidate antigens by cytotoxic T lymphocytes from Zebu cattle

East Coast fever (ECF) is a highly fatal lymphoproliferative disease of cattle caused by Theileria parva, a tick-borne intracellular apicomplexan parasite. Parasite antigens that are targets of protective cytotoxic T lymphocyte (CTL) responses are required to formulate a sub-unit vaccine against ECF. A number of CTL target antigens have recently been identified and initial evaluation has shown their vaccine potential. This study aimed to evaluate whether these antigens were recognised by CTL obtained from six genetically diverse Zebu cattle immunized with a cocktail of T. parva stocks. T. parva Muguga specific polyclonal CD8(+) CTL lines were generated and confirmed to specifically lyse autologous infected cells. CTL recognition of autologous skin fibroblasts (iSF) transduced with recombinant modified vaccinia virus Ankara strain (MVA) expressing previously identified T. parva Muguga vaccine candidate antigens was evaluated using an IFN-gamma ELISpot assay. CTL lines from one of the four calves, BY120, responded specifically to cells infected with MVA expressing the antigen Tp2 and synthetic peptides were employed to map a new CTL epitope on this antigen. Immunoscreening of the T. parva genome with these CTL lines should identify novel antigens that will constitute valuable additions to the vaccine candidates currently being evaluated.     

Comparison of indirect fluorescent antibody test and enzyme linked immunosorbent assay in the detection of exposure of cattle to Theileria parva in Kenya.

Appraisal of the indirect fluorescent antibody test (IFAT) and antigen enzyme linked immunosorbent assay (ELISA) serological tests as carried out to detect cattle exposed to Theileria parva at the National Veterinary Research Centre, Muguga (NVRC), Kenya is reported. Using sera from T. parva naive cattle and cattle experimentally exposed to T. parva, the two tests were appraised in terms of their sensitivity and specificity. IFAT and ELISA had the same sensitivity of 90% while ELISA had a higher specificity (90%) than IFAT (80%). A comparison was also made of the capability of the two tests to detect exposure of dairy cattle to T. parva prior to immunization against East Coast fever (ECF). The positive outcome from the IFAT was significantly higher (chi 2 = 30.36; P < 0.001) than that from the ELISA. The agreement between the two tests was low (Kappa = 0.21). The two tests indicated a higher risk of ECF in the study area than was expected. Indications are that the ELISA has been effectively adopted at NVRC.