Department of Biochemistry, Molecular Biology and Biotechnology
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Item Prevalence of canine gastrointestinal parasites in Morogoro, Tanzania(Taylor and Francis, 1996-07) Muhairwa, A. P; Mtambo, M. M.A; Kusiluka, L. J.M; Maeda, G. E; Kambarage, D. M; Makene, V. WOut of 235 domestic dogs from Morogoro municipality and Mgeta area in Morogoro region coproscopically screened for gastrointestinal parasitic infections, 174 (74%) were found positive for one or the other parasite. Ancylostonza caninunz was the most common parasite (72%). Protozoan parasites and other helminths were less prevalent (4%). The prevalence of gastrointestinal parasitism was significantly higher (P<0.05) in dogs <6 months (prkppies) than in adults with no sex effect.Item Intestinal protozoan parasites of pigs reared under different management systems in Morogoro, Tanzania(Taylor and Francis, 1996-07) Muhairwa, A. P; Mtambo, M. M.A; Kambarage, D. M; Esrony, K; Kusiluka, L. J.MSixty three piggery units with a total herd size of 424 pigs i n the small-scale and semi-intensive management systents from semi arid and tropical highlund areas of Morogoro region were screened for intestinal protozoan parasites. Thirtysix percent of the screened animals were positive for eimeriosis and 7.8% for cryptosporidiosis. Prevalences of eimeriosis in the semi-intensive and sntall-scale management systems were 22% and 48%, respectively. High prevalences of eimeriosis and cryptosporidiosis (60% and 25%, respectively) were observed in the tropical highland climate area (Mgeta) and low rates (26% and 0.3%, respectively) were evident i n the semi-arid zone. Ninety five per cent of infected pigs excreted between 100 and 5000 OPG and five per cent were excreting moreItem Eimeriosis in dairy cattle farms in Morogoro municipality of Tanzania(Elsevier Science B.V., 1996-10) Muhairwa, A.P; Chibunda, R.T; Kambarage, D.M; Mtambo, M.M.A; Kusiluka, L.J.M; Kazwala, R.RCoccidial oocysts were detected in 35% of 445 cattle in four medium-scale and 20 small-scale dairy farms in Morogoro municipality, Tanzania. The highest prevalence (56%) was observed in animals aged between 5 and 18 months, whereas lower prevalences were observed in calves (29%) aged between 12 days and 4 months and adults (30%). No coccidial oocysts were detected in calves less than 12 days old. The oocyst output was high in calves, followed by weaners; adults had the lowest oocyst output. The number of oocysts per gram of faeces was significantly higher (P < 0.001) in diarrhoeic animals than in non-diarrhoeic animals, and more so in young calves. Eimeria species infecting the animals included Eimeria bovis (68%) and Eimeria zuemii (57%), Eimeria ellipsoidalis (25%), Eimeria cylindrica (23%), Eimeria aubumensis (22%), Eimeria alabamensis (12%) and Eimeria subspherica (5%). Mixed infections involving two or three species were common. Our findings indicate that eimeriosis is common in cattle in Morogoro municipality. Published by Elsevier Science B.V.Item Helminthosis in local and cross-bred pigs in the Morogoro region of Tanzania(Elsevier, 1997-01) Muhairwa, A.P; Mtambo, M.M.A; Kambarage, D.M; Elsrony, K; Kusiluka, L. J. MWe investigated the prevalence, burden and types of gastro-intestinal helminths in 424 local and cross-bred pigs kept under different management systems in two climatic zones in the Morogoro region of Tanzania. Coprological examination revealed that 53% of the pigs excreted hehninth eggs in their faeces. The median eggs per gram of faeces (EPG) was 500 (range 100-22000). Local breeds in the Mgeta location with tropical highland climate showed signifi- cantly higher prevalence (P < 0.001) and median EPG values ( P < 0.001) than the cross-bred animals in the semi-arid area. There was no significant difference in the prevalence (P = 0.90) of helminth infection and egg outputs (P = 0.78) in cross-bred pigs raised under the small-scale and semi-mtensive management systems in the semi-arid zone. Piglets showed significantly lower prevalence of helminthosis (P < 0.001) than the weaners, growers and adults in both local and cross-bred animals. Median EPGs of growers and adult animals were significantly higher than those of piglets and weaners (P = 0.006). The prevalences of various hehninth species were Oesophagostomum sp. (40%), Ascaris suum (12%), Strongy loides ransomi (9% ) and Trichuris suis (St% ). 0 1997 Elsevier Science B.V.Item Investigations on the carrier rate of Pasteurella multocida in healthy commercial poultry flocks and flocks affected by fowl cholera(Taylor and Francis, 2000) Muhairwa, A. P; Christensen, J. P; Bisgaard, MTwenty flocks of web-footed birds (Pekin and Muscovy ducks and geese) and eight flocks of chickens raised under intensive management were examined for the presence of carriers of Pasteurella multocida. Five hundred and seventy-eight web-footed birds and 240 chickens from healthy flocks, as well as from flocks affected by fowl cholera, were investigated. A total of 135 isolates (80 from healthy flocks and 55 from flocks affected by fowl cholera) were obtained from the pharyngeal and cloacal mucosae after mouse passage (134 isolates) and culture in selective medium (one isolate). Thirty-five percent (7/20) of the flocks of web-footed birds and 38% (3/8) of chicken flocks had birds carrying P. multocida in the pharynx and/or cloaca. Birds from flocks affected by fowl cholera carried P. multocida at a significantly higher prevalence in the mucosa of the cloaca (P < 0.001) compared with the pharynx, while the opposite was observed in birds from healthy flocks. Extended phenotypic characterization confirmed the presence of P. multocida ssp. multocida, P. multocida ssp. septica and P. multocida ssp. gallicida in the flocks examined. P. multocida ssp. gallicida was exclusively isolated from Pekin ducks, while P. multocida ssp. multocida and P. multocida ssp. septica were obtained from chickens as well as web-footed birds. Each flock was shown to be infected by a single phenotypic clone, but some clones were found in more than one flock. A different clone was found in each of four outbreaks of fowl cholera on one of the farms in the preceding 2 years. Two genotypic and phenotypic clones each of P. multocida ssp. multocida and P. multocida ssp. septica were found. This observation indicated that outbreaks are usually clonal and that elimination of P. multocida from infected farms is possible. The results suggest that healthy poultry, in addition to convalescent carriers, may also be carriers of P. multocida. However, the virulence of P. multocida isolates and resistance of carriers to clinical infection needs to be examined. This is the first report of isolation of P. multocida from the cloacal mucosa of apparently healthy domestic poultry. Sampling of the cloaca appeared to be more sensitive for detecting carriers of P. multocida. Although selective medium was used only to a limited extent, the results suggested that mouse inoculation was a more efficient method of isolating P. multocida from poultry than the use of selective media.Item Occurrence of Pasteurella multocida and related species in village free ranging chickens and their animal contacts in Tanzania(Elsevier, 2000-07-13) Muhairwa, A.P.; Mtambo, M.M.A.; Christensen, J.P.; Bisgaard, M.Investigation was done to determine the presence of Pasteurella multocida and related species in free ranging chickens and ducks, dogs, cats and pigs in three climatic zones (cool, warm and hot) of rural Morogoro, Tanzania. A total of 153 isolates of P. multocida ssp. multocida and related species were obtained by direct culture on blood agar, selective medium and mouse inoculation. P. multocida ssp. multocida was isolated from 0.7% of chickens and 7% of ducks. In dogs and cats, P. multocida ssp. multocida was isolated from 1 and 68%, respectively. One isolate of Pasteurella gallinarum was isolated from a duck. Other species obtained were; P. multocida ssp. septica, Pasteurella stomatis and taxon 16 from dogs and cats, while Pasteurella dagmatis and Pasteurella canis were found in dogs only. Prevalence of P. multocida ssp. multocida was signi®cantly higher (P < 0:01) in ducks of the warm zone (22%) than in ducks of other zones (0%). No signi®cant difference was observed between the prevalence of P. multocida ssp. multocida in chickens of the warm zone (2%) and chickens of the cool and hot zones (0%). Extended phenotypic characterization revealed phenotypic similarities between two isolates from chickens and the duck strains. Mouse inoculation appeared to be more sensitive in detecting P. multocida ssp. multocida than blood agar and selective medium. Direct culture on blood agar recovered most of the isolates from dogs. This study has demonstrated for the ®rst time the presence of P. multocida and related species in the village free ranging chickens, ducks, dogs and cats in Tanzania. Other non-classi®ed Pasteurella spp. were also observed in the study, but further characterization is required before the ®nal classi®cation can be made. This paper reports for the ®rst time the isolation of unclassi®ed Pasteurella from dogs and cats in Africa. The results implies that fowl cholera might be occurringItem Serum resistance of Pasteurella multocida in avian and porcine sera, and comparative virulence investigations of selected serum-sensitive and resistant strains in chickens(Taylor and Francis, 2002) Muhairwa, Amandus P; Christensen, Jens P; Bisgaard, MagneGrowth in serum of Pasteurella multocida and related species in chicken, turkey, duck and pig sera were compared, and selected serum-resistant and serum-sensitive strains were inoculated into 18-week-old layers. Eighty-seven field strains of Pasteurella spp. and nine reference strains representing different clones defined by restriction endonuclease analysis ( REA) profiles were used in the study. Serum activity was measured by changes in the optical density ( OD) of the serum after inoculation and incubation at 41°C for chicken, turkey and duck serum and 39°C for pig serum. Serum activity was measured by comparison with previously determined serum-resistant ( P-1059) and serum- sensitive ( CU vaccine ) strains, and classified into highly serum-resistant, moderately serum-resistant and serum-sensitive. Strains of the same REA type were found to have identical growth curves and the same maximum OD values when tested in serum from the same host species. Turkey serum was shown to be less inhibitory to a wide range of P. multocida strains than chicken, duck and pig sera. Serum- resistant strains were demonstrated among avian as well as mammalian strains. Among the avian strains, the proportion of serum-resistant strains was higher in outbreak strains than in strains from apparently healthy carriers. Removal of the capsule from selected strains by hyaluronidase treatment failed to change the serum activity. The most severe lesions in experimentally infected chickens were produced by a serum-resistant strain; however, lesions were also found in chickens infected by serum- sensitive strains, indicating the involvement of multiple factors in the virulence of P. multocida. Further investigations on serum resistance are indicated in order to relate other host and bacterial factors responsible for the development of fowl cholera.Item The effect of concurrent infections with pasteurella multocida and ascaridia galli on free range chickens(Elsevier, 2002-01-26) Dahl, C.; Permin, A; Christensen, J.P; Bisgaard, M; Muhairwa, A.P; Petersen, K.M.D; Poulsen, J.S.D; Jensen, A.LPasteurella multocida and Ascaridia galli are observed with high prevalences in free range chickens in Denmark, but the impact is unknown. A study was carried out to examine the interaction between A. galli and P. multocida in chickens and the impact on production. Five groups, each with 20 18-week-old Lohmann Brown chickens were infected. Group 1 was orally infected with 1000 50 embryonated A. galli eggs. Group 2 received 10 4 cfu P. multocida intratracheally. Group 3 was infected with A. galli and subsequently with P. multocida. Group 4 was infected with P. multocida followed by A. galli. Group 5 was the control. The study ran for 11 weeks where clinical manifestations, weight gain and egg production were recorded. Excretion of P. multocida was determined on individual basis and blood smears were made for differential counts. At the end of the study pathological lesions and the number of adult worms, larvae and eggs in the faeces were recorded. The birds were more severely affected when infected with both pathogens compared to single infections with A. galli or P. multocida, respectively. A lower weight gain and egg production was observed with dual infections. A. galli infection followed by a secondary P. multocida infection resulted in more birds with pathological lesions and continued P. multocida excretion. In conclusion a negative interaction between A. galli and P. multocida was observed and it is postulated that free range chickens are at higher risk of being subjected to outbreaks of fowl cholera when they are infected with A. galli. # 2002 Elsevier Science B.V. All rights reserved.Item Influence of complementary food on growth and iron status of infants aged 6-12 months in Kilosa district Tanzania.(Sokoine University of Agriculture, 2004) Mamiro Peter Ruwaichi SimonChildhood malnutrition remains a common and major problem in Tanzania. Protein energy malnutrition (PEM) and micronutrient deficiencies are the major problems that j occur during the transitional phase from breast milk to solid complementary foods J (CFs) in infants. According to WHO (1999), PEM among under-fives in Tanzania stands at 31% while iron deficiency anemia affects 32% of the infants. Others include I'* iodine deficiency disorders (25%) and Vitamin A deficiency (6%). Most studies have ' associated inadequate intake and poor utilization of nutrients at the complementing age as the immediate causes ofthese problems. In the jfirst chapter various literature sources were consulted. General situation with regard >to malnutrition among children including some pertinent causative factors are I . discussed. A brief discussion on breastfeeding, CFs and micronutrient availability ’■ I ♦ from CFs is presented along with the importance of phytic acid in micronutrient availability. Various techniques that have been used to increase energy density ofCFs such as germination and fermentation are described. Quality and safety aspects ofCFs with regard to contamination with bacteria, cyanides and mycotoxins have been reviewed. These are important because they might be the potential sources of various diseases affecting childrens’ health. Production of low cost CFs that can be afforded by the majority of children, who are faced with nutritional problems, especially in the ruralareasofdevelopingcountriesisdiscussed. Finally,successandfailurestoriesof intervention programs that were implemented to solve nutritional problems among communities in developing countries from early senventies to the recent years community trials are presented.Item The evolutionary dynamics of genetic determinants of plasmodium falciparum resistance to sulfadoxine/pyrimethamine (sp) in South Eastern Tanzania(Sokoine University of Agriculture, 2008) Malisa, Allen LewisThis study reports a systematic follow up of genetic changes in the genes encoding the dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) enzymes of Plasmodium falciparum, in isolates from three rural districts of South-eastern Tanzania. The enzymes are the target of antimalarial drug, sulphadoxine- pyrimethamine (SP). The population-wide analysis of resistance mutations in the dhfr and dhps genes was applied to examine the influence of different drug use policies and their potential for the selection for SP resistance. A total of 47 244 bloodspot filter paper samples were collected from all individuals of all ages in randomly selected households in a series of annual surveys conducted between 2000 - 2006. Twenty percent (9 662) of all samples were found positive for P. falciparum infection on microscopy and hence were used for the genetic studies. DNA was extracted from P. falciparum-positxvz samples and dhfr and dhps genes were amplified by a nested polymerase chain reaction (PCR) and resistance conferring point mutations determined. Size polymorphisms at three sets of microsatellite loci linked to dhfr and three other sets of unlinked microsatellite loci were analysed by PCR amplification and electrophoresis on an automated sequencer. The influence of National treatment policy on the parasite reservoir was profound. The change of first line therapy from CQ to SP brought about highly significant increase of the frequencies of dhfr triple and dhps double mutants. Artemisinin-based combination therapy (ACT- SP+Artesunate) in Rufiji had a small and non-significant impact on the frequency of dhps double and dhfr triple mutant alleles, but significantly disrupted their association. Z)A/r-l inked microsatellites revealed high diversity around the dhfr sensitive alleles and significantly reduced diversity around mutant dhfr alleles. The majority of triple mutant alleles had one flanking microsatellite haplotype which has previously been shown to be derived from Southeast Asia, while the double mutant alleles had multiple haplotypes which were independently derived. Distribution of major lineages indicates that there is extensive genetic exchange among the geographic regions. Unlinked microsatellites confirmed the extent of allele sharing among the regions and revealed a major trend for reduced transmission intensity, which was apparently independent of the ACT intervention.Item Seroprevalence and factors affecting canine monocytic ehrlichiosis and canine brucellosis in Tanzania(Roavs, 2012) Muhairwa, A. P.; Msoffe, P. L. M.; Mtambo, M. M. A.; Mwakijungu, E. O.A cross-sectional study was undertaken to determine the seroprevalence of Ehrlichia canis and Brucella canis in dogs in Morogoro Tanzania. The study was conducted between June and September 2010. A total of 100 randomly selected dogs were tested for the presence of Ehrlichia canis and Brucella canis antibodies using the Immunocomb ® dot-ELISA tests (Biogal, Israel). Epidemiological factors such as age, sex, breed, health status, body condition and tick infestation were studied. E. canis antibodies were detected in 25% (n=100) of the dogs. B. canis antibodies were not detected in any of the study dogs. The difference in seroprevalence between old and adult dogs was statistically significant (P<0.05). There was also a significant difference in seroprevalence between dogs in good and those in fair body conditions (P<0.05). Seropositivity to E. canis was not associated with the other epidemiological factors. This study provides the first serological evidence of E. canis infection but found no evidence of antibodies to B. canis in dogs in Morogoro. Canine ehrlichiosis was found to be a prevalent disease in Morogoro and calls for regular testing and treatment of clinical cases and tick control measures to protect dogs from E. canis infection. The study also points out the need for further investigation on the presence of canine brucellosis.Item Awareness, knowledge and practice of pastoralists and agro- pastoralists towards livestock diseases affecting domestic animals in Arusha, Manyara and Morogoro Regions, Tanzania(2013) Chengula, A; Mdegela, R.H; Kasanga, C.JThe study was carried out to assess pastoralists and agro-pastoralists awareness, knowledge and practice in various livestock diseases affecting domestic animals in Arusha Manyara and Morogoro regions in Tanzania. Closed- and open-ended questionnaires, focus group discussions and in-depth interview techniques were employed. Diseases, drought, lack of dipping tanks, insufficient of livestock experts and drugs are the main constraints in the livestock keeping community in the study area. Nineteen diseases have been reported to affect their animals at one time or the other. East Coast fever (ECF, 79.7%), Contagious Caprine Pleuropneumonia (CCPP, 60.8%) and Trypanosomosis (50%) have been reported by more than 50% of pastoralists that they affect their animals. ECF and CCPP seem to be the leading diseases with great impact to the pastoralists by causing high mortality rates. Rift valley fever (RVF) and anthrax have been reported by majority to be diseases which appear in form of outbreak in their area. Diseases reported here are said to be controlled primarily by treating with various drugs and Oxytetracycline being a common drug of choice for most unknown diseases. Other control methods include vaccination and deworming, dipping and spray of animals using acariceides. Livestock experts have been reported to play little role in controlling common livestock diseases as majority of livestock keepers tends to treat their animals. Veterinary experts seem to be important during outbreak of diseases or for unknown diseases killing many animals. Eating of dead and improperly cooked meat together with un-boiled milk was found to be common in the pastoral community. This could lead to the spread (if any) of zoonotic diseases easily. Livestock keeping community is aware of most of common diseases circulating in their area but the way they practice to control leads to failure of control of those diseases at individual and national level. Devising a mechanism to educate them so that they know how to handle some common and reporting outbreak diseases such as use of trained community animal health workers (CAHWs) will help control livestock diseases in Tanzania.Item Rapid, sensitive and effective diagnostic tools for foot- and-mouth disease virus in Africa(2014) Kasanga, Christopher J; Yamazaki, Wataru; Mioulet, Valerie; King, Donald P; Mulumba, Misheck; Ranga, Ezekia; Deve, Jimis; Mundia, Cornelius; Chikungwa, Patrick; Joao, Laureta; Wambura, Philemo N; Rweyemamu, Mark MSpeed is paramount in the diagnosis of highly infectious diseases, such as foot-and-mouth disease (FMD), as well as for emerging diseases; however, simplicity is required if a test is to be deployed in the field. Recent developments in molecular biology have enabled the specific detection of FMD virus (FMDV) by reverse-transcription loop-mediated isothermal amplification (RT-LAMP), real-time reverse-transcription polymerase chain reaction (RT- qPCR) and sequencing. RT-LAMP enables amplification of the FMDV RNA-dependent RNA polymerase 3D(pol) gene at 63 °C (in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase) for 1 h, whilst RT-qPCR amplifies the same gene in approximately 2 h 30 min. In this study, we compared the sensitivity and effectiveness of RT-LAMP against RT-qPCR for the detection of the FMDV 3D(pol) gene in 179 oesophageal- pharyngeal scraping samples (collected by probang) obtained from clinically healthy cattle and buffalo in Malawi, Mozambique and Tanzania in 2010. The FMDV detection rate was higher with RT-LAMP (30.2%; n = 54) than with RT-qPCR (17.3%; n = 31). All samples positive by RT-qPCR (Cq ≤ 32.0) were also positive for the RT-LAMP assay; and both assays proved to be highly specific for the FMDV target sequence. In addition, the VP1 sequences of 10 viruses isolated from positive samples corresponded to the respective FMDV serotypes and genotypes. Our findings indicate that the performance of RT-LAMP is superior to RT-qPCR. Accordingly, we consider this test to have great potential with regard to the specific detection and surveillance of infectious diseases of humans and animals in resource-compromised developing countries.Item Determination of the presence of babesia DNA in blood samples of cattle, camel and sheep in Iran by PCR(Belgrade, 2014-10-13) Khamesipour, Faham; Doosti, Abbas; Koohi, Arman; Chehelgerdi, Mohammad; Mokhtari-Farsani, Abbas; Chengula, Augustino AlfredBabesia species are protozoan parasites that parasitize the erythrocytes of domestic animals and humans, caus- ing anemia in the host affected. These parasites cause a zoonotic disease known as babesiosis. Polymerase chain reaction (PCR) has proven to be very sensitive for detecting Babesia in blood samples of affected animals, particular in ruminants. The purpose of the current study was to determine the presence of Babesia DNA in the blood samples obtained from cattle, camel and sheep in Iran. In addition, the study aimed at establishing a rapid, reliable, specific and sensitive molecular tool, the PCR, for the detection of Babesia DNA in ruminants and dromedaries. Blood samples were collected from 372 rumi- nants and dromedaries (155 cattle, 95 sheep and 122 camel) kept at the Livestock Experimental Station. The animals came from randomly selected herds located in the important livestock-production regions of Iran of Isfahan and Chaharmahal va Bakhtiary during December 2012 to March 2013. PCR was used to detect Babesia DNA in the blood samples whereby an amplified band size of 428 bp was considered positive for Babesia spp. The results indicated that 7.10% (n= 155), 6.56% (n= 122) and 0.00% (n= 95) of the blood samples from cattle, camel and sheep were positive for Babesia DNA, respectively. The findings from this study revealed that there were Babesia DNA in blood taken from cattle and camel. To our knowl- edge, this is the first report to show the presence of Babesia DNA in blood samples of Iranian ruminants and dromedaries in Chaharmahal Va Bakhtiari and Isfahan provinces by PCR method. Though, diagnosis of low-level infections by the pa- rasite is important for the epidemiological studies. Our findings support the power of PCR test for Babesia DNA detection in blood samples and could be easily used for routine diagnosis.Item Assessing the awareness, knowledge, attitude and practice of the community towards solid waste disposal and identifying the threats and extent of bacteria in the solid waste disposal sites in Morogoro municipality in Tanzania(IISTE, 2015) Chengula, Augustino; Lucas, Bahati K; Mzula, AlexandaSolid wastes comprise all the wastes arising from human and animal activities that are normally solid, discarded as useless or unwanted materials. Health hazards associated with improper disposal of solid wastes to the community were investigated in Morogoro municipality. The aim of the project was to investigate the solid waste disposal practices and their health hazard implications to the community in Morogoro municipality. The study was conducted by considering several solid waste disposal sites based on three methods; observation, questionnaire survey and microbiological analysis. Based on observation method, several solid waste practices were detected including collection of wastes using trucks, wheel barrow, carriers made from elephant grasses and cement bags. Questionnaire survey pinpointed several diseases caused by solid wastes such as malaria, diarrhea, dysentery, cholera, typhoid and worm diseases from the respondents. From microbiological analysis, several pathogenic bacteria were identified from the solid disposal sites. The bacteria with their frequency of isolation identified were: Salmonella typhimurium (16.7%), Shigella dysenteriae (16.7%), Citrobacter freundii (8.3%), Citrobacter amalonaticus (8.3%), Aerobacter aerogenes (8.3%), Proteus vulgaris (16.7%), Klebsiella oxyotoca (8.3%), Klebsiella (8.3%), E.coli (8.3%). Solid waste generated by the daily activities of the people needs to be properly managed in such a way that it minimizes the risk to the environment and human health. Inadequate collection and disposal of solid waste is a major factor in the spread of disease and environmental degradation.Item Bioactivity of Synadenium glaucescens (pax) extracts on helminth eggs and larvae from wastewater stabilization ponds in Morogoro municipality,Tanzania(Sokoine University of Agriculture, 2015) Hassan, S.The increased use of wastewater and sludge from wastewater stabilization ponds in agriculture predisposes the consumers to the health risks. The objective of this study was to evaluate the bioactivity of Synadenium glaucescens (Pax) extracts on helminth eggs and larvae from wastewater and sludge in Morogoro. Helminth eggs and larvae were recovered using Bailenger and Baerman methods, and then quantified using McMaster techniques. Extracts (S. Glaucescens) used during hatchability and larvicidal bioassays were obtained using hot and cold solvent extraction. The effect of S. glaucescens extracts on hatchability and larvicidal was tested using in vitro and in vivo methods. One litre of wastewater was collected from anaerobic, facultative and maturation ponds. One hundred grams of sludge were sampled from the ponds and piles. Lethal concentration fifty (LC50) and inhibitory concentration fifty (IC50) were used to determine larvicidal and hatchability effects. The identified helminth eggs were from nematodes including Ascarid, Strongylid and Trichuris. Minimum amount of ≤ 1 e.p.l. were found in wastewater sampled from maturation ponds, same amount recommended by WHO guideline of 2006, and a maximum of 700 e.p.l. from anaerobic ponds. It was found that sludge samples contained a minimum of ≤ 1 e.p.g. from maturation ponds and maximum of 100 e.p.g. in anaerobic pond. The ethanol extracts of root bark and leaves were the most effective with IC50 19.34 and 39.56 μgml-1. The two extracts also demonstrated the highest LC50 of 19.41 and 30.19 μgml-1 respectively. The root bark extracts were more active than leaves extracts. This study demonstrated a high potential of using S. glaucescens extract in controlling helminths in wastewater and sludge.Item Molecular diagnosis and characterisation of orf virus in symptomatic goats in coast and Dar es salaam regions, Tanzania(Sokoine University of Agriculture, 2015) Mayenga, CharlesOrf virus (ORFV) is a member of the parapoxvirus genus that causes orf, a zoonotic and epitheliotropic highly contagious disease mainly affecting sheep, goats, wild ruminants and humans. In the present study, an outbreak of a disease in goats with clinical signs suggestive of orf in 11 flocks with a total of 259 goats was investigated between May and June 2015. Eight villages in districts of Bagamoyo (2), Ilala (1), Kinondoni (1) and Kisarawe (4) in Coast and Dar es Salaam regions of Tanzania were involved. The aim of the present study was to confirm ORFV involvement in diseased goats and to provide the genetic characteristics of ORFV. Upon visiting of goat flocks, a total of 72 goats presented orf-like clinical signs and 24 were reported to have died with similar clinical presentation. The presence of ORFV in oral swabs, scabs and skin scrapings was investigated by partial amplification of the ORFV RNA polymerase gene using Orf1 and Orf2 primers by polymerase chain reaction (PCR). A total of 16 out of 22 goats tested positive for ORFV upon PCR. Afterwards, molecular characterisation of ORFV was performed by amplification and nucleotide sequencing of the B2L gene encoding the major envelope protein. The results of nucleotide sequencing showed that orf was caused by closely related ORFV belonging to cluster I. ORFV were found to be genetically closely related to OV-SA00 (Accession number AY386264) strain of ORFV isolated in 2003 from scab material of a kid in United States of America and ORFV collected from a goat in Kyela, Tanzania in 2013. To our knowledge, this is the first report of the phylogenetic analysis of B2L gene of ORFV from goats in Tanzania. More studies are required to determine the extent of spread and genetic diversity of ORFV in livestock, wildlife and humans.Item Polymorphisms of Kappa-casein and Beta-lactoglobulin genes and their association with milk fat and protein contents in Tanzanian goats(Sokoine University of Agriculture, 2015) Kuwi, Salum OmariThis study was conducted to assess the polymorphisms of beta-lactoglobulin (-Lg) and kappa-casein (-Cn) genes and determine the effects of -Lg polymorphism on percentage of milk fat and protein contents in indigenous goat breeds found in Tanzania. Blood samples were collected from 22 - 24 unrelated animals for each breed. Genomic DNA was isolated from whole blood samples using Sepa gene extraction kit with some modifications. The -Lg and -Cn genes were amplified using Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) technique was used to determine variations at the -Lg and -Cn loci. The amplified products for -Lg and -Cn genes had the sizes of 426 and 459 bp, respectively. Digestion of β-Lg gene using SacII endonuclease restriction enzyme revealed the presence of two common alleles (A and B) in all goat populations. The frequency of B-allele (0.583) was higher than that of the A-allele (0.417) for β-Lg gene in indigenous goats, while exotic goats had higher frequency of A allele (0.688) than the B-allele (0.312). The frequencies of AA, AB and BB genotypes were, respectively, 0.250, 0.333 and 0.417 in Gogo goats, 0.318, 0.273 and 0.409 in Pare white, 0.417, 0.333 and 0.250 in Malya, 0.500, 0.375 and 0.125 in Saanen. The analysis of -Cn gene variation in the four goat populations indicated that there were three alleles (A, B and C) at this locus. The observed allelic frequencies of A/B and C were 0.917 and 0.083 in Gogo white, 0.955 and 0.045 in Pare white, 1 and 0.00 in Malya, 0.875 and 0.125 in Saanen goats, respectively. The effects of β-Lg genotypes on milk fat and protein percentages were analyzed using a general linear model (GLM). The results show that there was no significant effects (P>0.05) of β-Lg alleles on milk fat and protein percentages.Item Sero-surveillance, risk factors and molecular diagnosis of peste des petits ruminants virus in South Kivu, Democratic Republic of Congo(Sokoine University of Agriculture, 2015) Ahadi, Bwihangane BirindwaPeste des petits ruminants (PPR) is an acute viral disease of small ruminants caused by PPR virus (PPRV). For many years, the disease was mainly confined to West and Central Africa but has now spread southwards to previously PPR-free countries. The disease was first reported in Democratic Republic of Congo (DRC) in 2012. The disease causes high morbidities of up to 100 % and mortality rates between 50 and 90% in domestic small ruminants (goats and sheep) leading to the socio economic loss impact to the farmers. This study aimed to perform molecular diagnosis and sero-prevalence of PPR associated with transmission risk factors in unvaccinated sheep and goats from South-Kivu in province in DRC using respectively reverse- transcriptase polymerase chain reaction (RT-PCR), competitive enzyme-linked immunosorbent assay (cELISA) and a structured questionnaire. The results showed with cELISA an overall seroprevalence of 28.5% (n=319), out of which 11.3% and 32.7% seropositivity was found in sheep and goats, respectively. Peste des petits ruminants seroprevalence was higher in the territories that recorded high rainfall, 34.5% (n=142) in Shabunda and 29.4% (n=79) in Mwenga. In a total of 11 risk factors investigated four were found to be associated with PPR seroprevalence (p≤0.05). Among them we have animal’s age (OR: 9.34), grazing and farming system (6.28), territory geographic location (OR: 5.1) and the animal’s origin (OR: 0.5). Peste de petits ruminants seroprevalence was higher in small ruminants kept in communal grazing system (30.6%) and free ranging system (31.2%). Sheep and goats of >12 months had a significantly higher PPR seroprevalence (35.1%). As no PPRV RNA was detected in any of the blood collected using reverse transcriptase polymerase chain reaction (RT-PCR), we recommend further studies to be focused on molecular characterization and isolation of PPR virus.Item Detection of Arenaviruses in rodents, shrews and elephant shrews from selected wildlife-human interfaces in Tanzania(Sokoine University of Agriculture, 2015) Maganga, RuthThe present study was conducted to investigate the presence of Arenaviruses from rodents, shrews and elephant shrews captured in selected wildlife-human interfaces in Tanzania. The study involved six sites with high potential for contact between wildlife and humans namely; Ruaha, Kilombero, Mtwara, Mbeya, Mbinga and Mikumi. A total of 121 animals comprising 111 rodents, 3 shrews and 7 elephant shrews were screened for Arenaviruses using conventional Polymerase Chain Reaction (PCR). The genetic relatedness of Arenaviruses was evaluated by conducting phylogenetic analysis of partial sequences of the S gene. The association between age and sex with the presence of Arenaviruses was assessed. Of the 121 animals, 7 (5.8 %) were shedding Arenavirus. All positive animal samples were obtained from the Ruaha site at crop raiding and peridomestic interfaces. Eighty six percent of the infected animals were Mastomys sp. and 14 % were Arvicanthis sp. Age and sex of the animals were not significantly associated with occurrence of Arenaviruses in rodents, shrews and elephant shrews (P > 0.05). Additionally, Arenavirus detection in fecal specimens was not significantly different from the detection in oropharyngeal specimens (P > 0.05), clearly indicating that both specimens are useful for Arenavirus testing. Phylogenetic analysis showed that isolates obtained from this study were related to the Old World Arenaviruses, and include strains of Morogoro virus and a novel strain of Arenavirus. In conclusion, the present study has confirmed the presence of Arenaviruses closely related to other known Old World Arenaviruses in the Ruaha ecosystem.