Molecular prevalence of trypanosome infections in cattle and tsetse flies in the Maasai Steppe, northern Tanzania.

African trypanosomosis is a disease of public health and economic importance that poses a major threat to the livelihoods of people living in the Maasai Steppe, where there is a significant interaction between people, livestock and wildlife. The vulnerability of the Maasai people to the disease is enhanced by the interaction of their cattle, which act as vehicles for trypanosomes, and tsetse flies close to wildlife in protected areas. This study was aimed at identification of trypanosome infections circulating in cattle and tsetse flies in order to understand their distribution and prevalence in livestock/wildlife interface areas in the Maasai Steppe. A total of 1002 cattle and 886 tsetse flies were sampled from June 2015 to February 2016 in five villages and PCR was conducted to amplify the internal transcribed spacer 1 (ITS1) from trypanosomes. All Trypanosoma brucei-positive samples were further tested for the presence of the serum resistance-associated (SRA) gene found in human-infective trypanosomes using the SRA-LAMP technique. The overall prevalence of trypanosome infections was 17.2% in cattle and 3.4% in tsetse flies. Using a nested PCR, prevalence and abundance of five trypanosome species, Trypanosoma vivax, T. brucei, T. simiae, T. theileri and T. congolense, were determined, which varied with season and location. The highest prevalence of the identified trypanosome species was recorded at the end of wet season with an exception of T. brucei which was high at the beginning of the wet season. No human-infective trypanosomes were detected in both cattle and tsetse fly DNA. This study confirms that seasonality and location have a significant contribution to the prevalence of trypanosome species in both mammalian and vector hosts. These results are important for designing of community-wide vector and disease control interventions and planning of sustainable regimes for reduction of the burden of trypanosomosis in endemic pastoral areas, such as the Maasai Steppe in northern Tanzania.

Simwango M ，Ngonyoka A ，Nnko HJ ，Salekwa LP ，Ole-Neselle M ，Kimera SI ，Gwakisa PS ... -  《Parasites & Vectors》

Occurrence of trypanosome infections in cattle in relation to season, livestock movement and management practices of Maasai pastoralists in Northern Tanzania.

African animal trypanosomosis (AAT) is a parasitic disease considered to be one of the greatest constraints to cattle production in Tanzania. There is insufficient information on seasonal occurrence of AAT and management practices in Monduli District of the Maasai Steppe ecosystem to guide and prioritize AAT control programs. A cross-sectional survey was undertaken in 10 randomly selected villages of Monduli District. Information on seasonal animal movements, including wildlife interaction, and AAT management practices was gathered using a standardized questionnaire with 130 pastoralists. Blood samples were collected from a total of 960 cattle, in wet and dry seasons. An entomological survey was also undertaken in the dry season. Polymerase chain reaction targeting the internal transcribed spacer 1 (ITS1) was used for parasite identification in cattle blood and in tsetse flies. The overall apparent prevalence of AAT in cattle was 5.8% (95% CI of 4.1-8.3) and 4.2% (95% CI of 2.7-6.3) during wet and dry reasons, respectively. Trypanosoma vivax was the most common species identified in cattle. All tsetse flies (n = 426) collected were trapped in Esilalei village which is in close proximity to Lake Manyara National Park. Tsetse fly infection status was determined to be 7.0%; (CI 95% of 4.9-9.8%) with nearly 50% of infections due to T. congolense. All 130 cattle owners reported that they could easily recognize AAT and the majority (75%) identified the most prominent clinical signs. Nearly all owners (98.5%) identified that tsetse flies were responsible for AAT transmission. All cattle owners (100%) reported the use of trypanocides for AAT treatment, while 2.3% reported to also use herbal medicine. The trypanocides, Novidium® and Berenil®, were the most frequently reported commercial drugs and were used by 42% of cattle owners. Vector control by hand spraying was reported by the majority (90.8%) of cattle owners, while dipping and deployment of insecticide-impregnated targets were reported by few cattle owners (16.2% and 5.4%, respectively). The majority of cattle owners (83.1%) reported to move cattle away from home villages during the dry season with many migrating to areas in close proximity to wildlife parks. This study confirms the presence of circulating pathogenic trypanosomes in tsetse flies which continue to pose a threat to Maasai cattle. The seasonal movement of cattle during the dry season was associated with more clinical cases of cattle trypanosomosis. This study demonstrates the need to strengthen surveillance and control strategies for AAT.

Kimaro EG ，Toribio JLML ，Gwakisa P ，Mor SM ... -  《-》

The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve.

Trypanosomes are protozoan flagellates that cause human African trypanosomiasis (HAT) and African animal trypanosomiasis (AAT). HAT is caused by e in East and Central Africa and in West Africa, whereas AAT is caused by a number of trypanosome species, including and The aim of this study was to establish if tsetse flies at Liwonde Wild Life Reserve (LWLR) are infected with these trypanosomes and thus pose a risk to both humans and animals within and surrounding the LWLR. A total of 150 tsetse flies were caught. Of these, 82 remained alive after capture and were dissected such that the mid-gut could be examined microscopically for trypanosomes. DNA extractions were performed from both mid-guts and the 68 dead flies using a Qiagen Kit. Amplification techniques involved the Internal Transcriber Spacer 1 (ITS 1) conventional polymerase chain reaction (PCR) with primers designed to identify trypanosome species, and Repetitive Insertion Mobile Element - Loop Mediated Isothermal Amplification (RIME LAMP), a sequence specific to T. brucei. Analysis showed that 79/82 (96.3%) of the mid-guts examined microscopically were positive for trypanosomes and that 75/150 (50%) of the DNA extracts (from the mid-gut, and tsetse fly carcasses) were positive for T. brucei, as determined by the RIME LAMP method. ITS1 PCR further showed that 87/150 (58.0%) flies were positive for trypanosomes, of which 56/87 (64.4%) were 9/87 (10.3%) were 7/87 (8.1%) were ; 6/87 (6.9%) were and 6/87 (6.9%) were Ten samples had a mixture of infections. Our analysis demonstrated a mixture of infections from trypanosome species in tsetse flies at LWLR, and that the species that causes HAT, was the most common. Our study successfully used molecular techniques to demonstrate the presence of at LWLR, a species that causes HAT in both East and Central Africa.

Nayupe SF ，Simwela NV ，Kamanga PM ，Chisi JE ，Senga E ，Musaya J ，Maganga E ... -  《-》

Polymerase chain reaction identification of Trypanosoma brucei rhodesiense in wild tsetse flies from Nkhotakota Wildlife Reserve, Malawi.

Array A polymerase chain reaction (PCR) technique was used to identify the serum resistance associated () gene of in tsetse flies. Of 257 tsetse flies that were randomly caught, 42 flies were dissected for microscopic examination. The midguts of 206 flies were positive and were individually put in eppendorf tubes containing phosphate-buffered saline (PBS buffer) for DNA extraction. Internal transcribed spacer (ITS)-PCR was first used to isolate all trypanosome species from the flies. TBR PCR was then used to isolate the group. -positive samples were further evaluated by SRA PCR for the presence of the gene. Of 257 flies caught, 185 (72%) were and 72 (28%) were . Three were tenerals and 242 were mature live flies. Of the 242 flies dissected, 206 were positive, representing an 85.1% infection rate. From 206 infected flies, 106 (51.5%) were positive using ITS-PCR, 68 (33.0%) being mixed infections, 18 (8.7%) 9 (4.4%) , 4 (1.9%) , 3 (1.5%) , 3 (1.5%) , and 1 (0.4%) . When subjected to TBR PCR, 107(51.9%) were positive for . Of the 107 positive samples, 5 (4.7%) were found to have the gene. These results suggest that wild tsetse flies in Malawi are infected with human-infective trypanosomes that put communities around wildlife reserves at risk of human African trypanosomiasis outbreaks. Further studies need to be done to identify sources of blood meals for the flies and for surveillance of communities around wildlife reserves.

Musaya J ，Chisi J ，Senga E ，Nambala P ，Maganga E ，Matovu E ，Enyaru J ... -  《-》

Seasonal variation of tsetse fly species abundance and prevalence of trypanosomes in the Maasai Steppe, Tanzania.

Tsetse flies, the vectors of trypanosomiasis, represent a threat to public health and economy in sub-Saharan Africa. Despite these concerns, information on temporal and spatial dynamics of tsetse and trypanosomes remain limited and may be a reason that control strategies are less effective. The current study assessed the temporal variation of the relative abundance of tsetse fly species and trypanosome prevalence in relation to climate in the Maasai Steppe of Tanzania in 2014-2015. Tsetse flies were captured using odor-baited Epsilon traps deployed in ten sites selected through random subsampling of the major vegetation types in the area. Fly species were identified morphologically and trypanosome species classified using PCR. The climate dataset was acquired from the African Flood and Drought Monitor repository. Three species of tsetse flies were identified: G. swynnertoni (70.8%), G. m. morsitans (23.4%), and G.pallidipes (5.8%). All species showed monthly changes in abundance with most of the flies collected in July. The relative abundance of G. m. morsitans and G. swynnertoni was negatively correlated with maximum and minimum temperature, respectively. Three trypanosome species were recorded: T. vivax (82.1%), T. brucei (8.93%), and T. congolense (3.57%). The peak of trypanosome infections in the flies was found in October and was three months after the tsetse abundance peak; prevalence was negatively correlated with tsetse abundance. A strong positive relationship was found between trypanosome prevalence and temperature. In conclusion, we find that trypanosome prevalence is dependent on fly availability, and temperature drives both tsetse fly relative abundance and trypanosome prevalence.

Nnko HJ ，Ngonyoka A ，Salekwa L ，Estes AB ，Hudson PJ ，Gwakisa PS ，Cattadori IM ... -  《-》