\ud Detection and Monitoring of Insecticide Resistance in Malaria Vectors in Tanzania Mainland\ud

\ud Vector control is a major component of the global strategy for malaria control which aims to prevent parasite transmission mainly through interventions targeting adult Anopheline vectors. Insecticide treated nets (ITNs) and indoor residual spraying (IRS) are the cornerstone of malaria vector control programmes. These major interventions in most cases use pyrethroid insecticides which are also used for agricultural purposes. With widespread development of resistance to pyrethroid insecticides in malaria vectors raises concern over the sustainability of insecticide-based interventions for malaria control. Therefore, close monitoring of performance of the insecticides against malaria vectors is essential for early detection and\ud management of resistance. To measure pyrethroid susceptibility in populations of malaria vectors in Tanzania and to test the efficacy of LLINs/ITNs and insecticide residues on sprayed wall substrates in the IRS operation areas. In 2011 the National Institute for Medical Research (NIMR) in collaboration with National Malaria Control Programme (NMCP) conducted large scale surveillance to determine the countrywide susceptibility levels of malaria vectors to insecticides used for both public health and agricultural purposes. Anopheles gambiae Giles s.l. were collected during national surveys and samples of LLINs/ITNs in the 14 sentinel sites and houses from the IRS areas were randomly selected for bioassays to test the efficacy and insecticide residual effects on sprayed wall substrates respectively. Wild adult mosquitoes for susceptibility testing were collected by resting catches indoors. Net traps (outdoors and indoors) were set up to enhance catches. WHO Susceptibility kits were used to test for resistance status using test papers: Lambdacyhalothrin 0.05%, Deltamethrin 0.05%, Permethrin 0.75%, DDT 4%, Propoxur 0.1% and Fenitrothion 1%. The quality of the test paper was checked against a laboratory susceptible An. gambiae Kisumu strain. Knockdown effect and mortality were measured in standard WHO susceptibility tests and cone bio-efficacy tests. Whereas, con bioassays on treated walls and ITNs were conducted using the laboratory susceptible An. gambiae Kisumu strain. The results from the surveillance recorded continued susceptibility of malaria vectors to commonly used insecticides. However, there were some isolated cases of resistance and/or reduced susceptibility to pyrethroid insecticides which may not compromise the current vector control interventions in the country. Anopheles gambiae s.l. showed resistance (15-28%) to each of the pyrethroids and to DDT but not to Organophosphates (Propoxur 0.1%), and Carbamates (Fenitrothion 1%). The information obtained from this surveillance is expected to be used to guide the National Malaria Control Programme on the rational selection of insecticides for malaria vector control and for the national mitigation plans for management and containment of malaria vector resistance in the country. The current observation warrants more vigilant monitoring of the susceptibility of malaria mosquitoes to commonly used insecticides in areas found with resistance and/or reduced levels of susceptibility of malaria vectors to insecticides, particularly in areas with heavy agricultural and/or public health use of insecticides where resistance is likely to develop. The current survey covered malaria vectors only and not the non malaria vectors (nuisance) mosquitoes such as Culex. Similar monitoring of insecticide susceptibility of this non malaria vectors may be needed to ensure public motivation for sustained use of ITNs/LLINs in the country. The surveillance leading to these results received funding from PMI/USAID through RTI International with Sub Agreement Number 33300212555.\u

Mosquito Net Coverage and Utilisation for Malaria\ud Control in Tanzania\ud

\ud In recent years malaria parasites have developed resistance to the most commonly used antimalarial drugs in Tanzania, posing a major challenge for its control. This has led to frequent changes of malaria treatment guidelines, more recently to expensive, yet more effective arthemether/lumefantrine. The use of insecticide treated mosquito nets (ITNs) and long lasting nets (LLINs) in Tanzania has increased slowly over the past few years. Despite the introduction of a voucher scheme to the vulnerable groups, the proportion of households with at least an ITN/LLIN in the country has not been able to achieve the Abuja Declaration of 60% net coverage. Statistics available on the utilisation of nets do not provide a good estimate of the coverage, because of the different study design used to collect the information. This survey was carried out in 21 districts of Tanzania to determine the coverage and utilisation of insecticide treated nets to provide baseline information of the net requirement to cover every sleeping bed in the country. Specifically, this study aimed to (i) determine the ITN coverage by and its distribution in the country; and (ii) determine knowledge, attitudes and practice of the householders as regards to malaria prevention and control Twenty one districts (one from each region) of Mainland Tanzania were selected for the survey. Selection of the district was random. In each district, two wards were selected, one urban (within the district capital) and one rural or sub‐urban. Households were selected randomly using a table of random numbers. At household level, the head or any adult who represented the head of household was interviewed. A structured pre‐tested questionnaire was used to collect information on knowledge, attitude and practices in malaria control, with emphasis on mosquito net ownership and utilisation. Of the 9549 targeted households, 9166 (96%) participated in the survey. Majority of the respondents (76.8%) were from rural district.The mean household size was 3.9 persons. On average, children <5 years old accounted for 39.3% of the members of the households. Respondents with no formal education accounted for 15.8‐37.4% of the interviewees. Most of them were from Mkuranga (55%), Kigoma‐Ujiji (44.2%) and Newala (37.9%). High literate rates were observed in Arumeru and Moshi districts. The majority of the respondents knew that the mosquito is the vector of the malaria parasite (92.6‐99.4%) and infection is through a mosquito bite (92.7‐99.8%). The knowledge of respondents on malaria transmission was generally high (94.0‐99.0%). The majority of the respondents (95.2%) considered the use of mosquito nets as the most effective way of malaria prevention. However, of these, only 66.7% said to have actually used nets in their life time. Knowledge on the use of mosquito nets in the control of malaria was highest and lowest in Eastern and Central zones, respectively.\ud Seventy‐seven percent (4457/8933) agreed to have the investigator entry into their houses and verify the\ud number of nets owned. On average, 62.9% (5,785/8933) of the households had at least a mosquito net. Majority of the respondents in Northern (76.5%) and Southern (76.5%) zones owned at least a mosquito net. The lowest mosquito net ownership was observed among respondents in Western Zone (39.6%). District‐wise, net ownership was highest in Lindi (94.5%), Kyela (91.3%) Arumeru (86.1%), Ilala (83.1%)\ud and Nyamagana (80.0%). Ownership of net was very low in Kilolo (34.8%), Kigoma (36.5%) and Musoma Rural (41.3%). Of the households with nets, 74.4% were using nets all year round. A larger proportion of respondents in Kilolo (68.5%), Mpwapwa (51.9%), Songea Rural (49.2%) and Shinyanga Rural (46.3%) were only using the nets during the rainy season. Out of 9,166 households visited, 3,610 (39.3%) had at least one under five child. Of these, in 1,939 (53.7%) of the households the child slept under a mosquito net during the previous night. Use of nets in children <5 years was most common in northern zone (74%); followed by eastern (66.9%) and southern zone (61.1%). Districts with the largest proportion of <5 year children sleeping under a mosquito nets were Lindi (90.0%), Kyela (85.2%), Ilala (83.2%) and Arumeru (78.2%). Only about a quarter (27%) of the children <5% in western zone were sleeping under a mosquito net. Lowest net coverage for <5 year was in Kigoma (22.7%), Kilolo (25%) and Bukoba Rural (31.2%). A total of 5,785 (62.9%) owned at least a mosquito net. Of these, 4,219 and 1,566 were from the rural and urban districts, respectively. More households in the urban districts (73.4%) than rural districts (59.7%) owned at least a mosquito net. Likewise, there were more households (64.9%) in the urban districts with <5years children sleeping under mosquito nets than in the rural districts (50.4%). More households in urban (32.8%) than in rural districts (25.1%) had at least one insecticide treated net. The number of households with mosquito nets enough for all members of the households ranged from 18.9% (in Urambo) to 37.4% (in Hanang). Households with at least 50% or more occupants using mosquito nets ranged from 16.4% (in Urambo) to 42.8% (in Arumeru). Districts with the largest proportion of ≥50% of the household members sleeping under mosquito nets were Arumeru (46.9%) and Lindi (46.7%). In Manyoni and Lindi, 3.1% and 5% of the households were found to have more nets than the number of household occupants. Only 9% (801/9196) of the households had all occupants sleeping under a mosquito net. Kyela district had about a quarter (23.9%) of the households with all occupants sleeping under nets. Only 29% of the households had at least one insecticide treated mosquito nets. All nets in 51.4% of the households surveyed were ITNs. The largest proportion of households with ITN was observed in northern zone (40.2%), with Arumeru (46%) and Hanang (44.1%) districts having the highest ITN coverage. The lowest proportion (15.5%) of households with ITN was found in the Western Zone. Districts which had the lowest ITN coverage were Musoma Rural (12.6%), Kigoma‐Ujiji (13.2%), and Shinyanga Rural (14.4%). On average, 90.7% (8,123/8,953) of the respondents would prefer using ITN than having their house sprayed with long lasting residual insecticide. More households in urban (32.8%) than in rural districts (25.1%) had at least an ITN> A total of 1939 children underfives were sleeping under mosquito net (any type). Of these, 1140 (58.8%) were using insecticide treated nets (ITN). Overall, 31.6% of the underfives slept under an insecticide treated net during the previous night. Highest coverage was reported in Kyela (47.7%), Nyamagana (47.7%) and Arumeru (46.4%). Lowest ITN in underfives was reported in Kigoma‐Ujiji (16.0%), Musoma (17.2%) and Urambo (17.7%). In Songea more underfives children were sleeping under ITN (43.6%) than in untreated nets (40.9%). Control of bedbugs, lice, fleas, mites and cockroaches was the major added advantage of using insecticide treated nets. On average, 30.8% and 19.6% of the respondents mentioned cockroach and bedbug control as the main advantage of using ITN, respectively. The majority (52.9%) preferred blue coloured net (Northern=45.6%; Central=59.2%; Eastern=56.4%; Lake= 54.4%; Southern= 60.3%, Western= 58.5%) and Southern Highlands= 49.1%). Other colour preferences were white (29.6%), green (14.1%), black (2.1%) and pink (1.2%). A strong preference for blue mosquito nets was observed among respondents in Musoma (77.3%) and Newala (75.5%) districts. On the other hand, the weakest preference (24.7%) for blue nets was observed among respondents in Arumeru district. The majority of the respondents (82%) preferred rectangular shaped net. A larger percent (61.8%) the respondents preferred to have the map of Tanzania as a national logo to identify nets distributed in the country. On average, 62.7% and 28.8% of the households in Tanzania own at least one mosquito net (any type) and insecticide treated net, respectively. Tanzania expects that ITN coverage of under fives in 2009, after the Under Five Catch‐up Campaign is complete, to be at least 80%. If this is to be achieved, there is a need for concerted effort in scaling up the distribution and demand for long lasting nets throughout the country. Moreover, the planned use of IRS in malaria control, currently considered unpopular should be accompanied by rigorous community health education to avoid resistance from household members.\u

Tumour-associated immune responses and isolated carcinoembryonic antigen and alpha feto-protein levels related to survival in ovarian cancer patients.

The presence of a tumour-associated immune response in 37 patients with ovarian cancer as assessed by blastogenesis (lymphocyte transformation) evoked by ovarian cancer cell extracts, has been correlated with survival following the test. The difference in these responses is unlikely to be accounted for on the basis of general impairment of cell-mediated immuno-competence. Serum carcinoembryonic antigen (CEA) was also determined in 27 ovarian cancer patients to assess its prognostic significance. Raised CEA levels and absence of blastogenic response to tumour cell extract during relapse are associated with a worse prognosis but neither of these parameters are significant in remission. Possible applications of these findings to the clinical management of ovarian cancer patients are discussed. Serum alpha feto-protein levels measured by radioimmunoassay were not found to be raised in any of the 32 ovarian cancer patients in whom it was measured

Causes of death and associated factors over a decade of follow-up in a cohort of people living with HIV in rural Tanzania

BACKGROUND: Nearly half of HIV-related deaths occur in East and Southern Africa, yet data on causes of death (COD) are scarce. We determined COD and associated factors among people living with HIV (PLHIV) in rural Tanzania. METHODS: PLHIV attending the Chronic Diseases Clinic of Ifakara, Morogoro are invited to enrol in the Kilombero and Ulanga Antiretroviral Cohort (KIULARCO). Among adults (>/= 15 years) enrolled in 2005-2018, with follow-up through April 2019, we classified COD in comprehensive classes and as HIV- or non-HIV-related. In the subset of participants enrolled in 2013-2018 (when data were more complete), we assessed cause-specific mortality using cumulative incidences, and associated factors using proportional hazards models. RESULTS: Among 9871 adults (65% female, 26% CD4 count < 100 cells/mm(3)), 926 (9%) died, among whom COD were available for 474 (51%), with missing COD mainly in earlier years. The most common COD were tuberculosis (N = 127, 27%), non-AIDS-related infections (N = 72, 15%), and other AIDS-related infections (N = 59, 12%). Cardiovascular and renal deaths emerged as important COD in later calendar years, with 27% of deaths in 2018 attributable to cardiovascular causes. Most deaths (51%) occurred within the first six months following enrolment. Among 3956 participants enrolled in 2013-2018 (N = 203 deaths, 200 with COD ascertained), tuberculosis persisted as the most common COD (25%), but substantial proportions of deaths from six months after enrolment onwards were attributable to renal (14%), non-AIDS-related infections (13%), other AIDS-related infections (10%) and cardiovascular (10%) causes. Factors associated with higher HIV-related mortality were sex, younger age, living in Ifakara town, HIV status disclosure, hospitalisation, not being underweight, lower CD4 count, advanced WHO stage, and gaps in care. Factors associated with higher non-HIV-related mortality included not having an HIV-positive partner, lower CD4 count, advanced WHO stage, and gaps in care. CONCLUSION: Incidence of HIV-related mortality was higher than that of non-HIV-related mortality, even in more recent years, likely due to late presentation. Tuberculosis was the leading specific COD identified, particularly soon after enrolment, while in later calendar years cardiovascular and renal causes emerged as important, emphasising the need for improved screening and management

Stigma-directed services (Stig2Health) to improve 'linkage to care' for people living with HIV in rural Tanzania: study protocol for a nested pre-post implementation study within the Kilombero and Ulanga Antiretroviral Cohort

Background: HIV-related stigma is a major barrier to the timely linkage and retention of patients in HIV care in sub-Saharan Africa, where most people living with HIV/AIDS reside. In this implementation study we aim to evaluate the effect of stigma-directed services on linkage to care and other health outcomes in newly diagnosed HIV-positive patients. Methods: In a nested project of the Kilombero and Ulanga Antiretroviral Cohort in rural Tanzania, we conduct a prospective observational pre-post study to assess the impact of a bundle of stigma-directed services for newly diagnosed HIV positive patients. Stigma-directed services, delivered by a lay person living with HIV, are i) post-test counseling, ii) post-test video-assisted teaching, iii) group support therapy and group health education, and iv) mobile health. Patients receiving stigma services (enrolled from 1 (st) February 2020 to 31 (st) August 2021) are compared to a historical control receiving the standard of care (enrolled from 1 (st) July 2017 to 1 (st) February 2019). The primary outcome is 'linkage to care'. Secondary endpoints are retention in care, viral suppression, death and clinical failure at 6-12 months (up to 31 (st) August 2022). Self-reported stigma and depression are assessed using the Berger Stigma scale and the PHQ-9 questionnaire, respectively. The sample size calculation was based on cohort data from 2018. Assuming a pre-intervention cohort of 511 newly diagnosed adults of whom 346 (68%) were in care and on antiretroviral treatment (ART) at 2 months, a 10% increase in linkage (from 70 to 80%), a two-sided type I error rate of 5%, and 90% power, 321 adults are required for the post-implementation group. Discussion: We expect that integration of stigma-directed services leads to an increase of proportions of patients in care and on ART. The findings will provide guidance on how to integrate stigma-directed services into routine care in rural sub-Saharan Africa

Predicting Distribution of Aedes Aegypti and Culex Pipiens Complex, Potential Vectors of Rift Valley Fever Virus in Relation to Disease Epidemics in East Africa.

The East African region has experienced several Rift Valley fever (RVF) outbreaks since the 1930s. The objective of this study was to identify distributions of potential disease vectors in relation to disease epidemics. Understanding disease vector potential distributions is a major concern for disease transmission dynamics. DIVERSE ECOLOGICAL NICHE MODELLING TECHNIQUES HAVE BEEN DEVELOPED FOR THIS PURPOSE: we present a maximum entropy (Maxent) approach for estimating distributions of potential RVF vectors in un-sampled areas in East Africa. We modelled the distribution of two species of mosquitoes (Aedes aegypti and Culex pipiens complex) responsible for potential maintenance and amplification of the virus, respectively. Predicted distributions of environmentally suitable areas in East Africa were based on the presence-only occurrence data derived from our entomological study in Ngorongoro District in northern Tanzania. Our model predicted potential suitable areas with high success rates of 90.9% for A. aegypti and 91.6% for C. pipiens complex. Model performance was statistically significantly better than random for both species. Most suitable sites for the two vectors were predicted in central and northwestern Tanzania with previous disease epidemics. Other important risk areas include western Lake Victoria, northern parts of Lake Malawi, and the Rift Valley region of Kenya. Findings from this study show distributions of vectors had biological and epidemiological significance in relation to disease outbreak hotspots, and hence provide guidance for the selection of sampling areas for RVF vectors during inter-epidemic periods

Mathematical Evaluation of Community Level Impact of Combining Bed Nets and Indoor Residual Spraying upon Malaria Transmission in Areas where the main Vectors are Anopheles Arabiensis Mosquitoes.

Indoor residual insecticide spraying (IRS) and long-lasting insecticide treated nets (LLINs) are commonly used together even though evidence that such combinations confer greater protection against malaria than either method alone is inconsistent. A deterministic model of mosquito life cycle processes was adapted to allow parameterization with results from experimental hut trials of various combinations of untreated nets or LLINs (Olyset, PermaNet 2.0, Icon Life nets) with IRS (pirimiphos methyl, lambda cyhalothrin, DDT), in a setting where vector populations are dominated by Anopheles arabiensis, so that community level impact upon malaria transmission at high coverage could be predicted. Intact untreated nets alone provide equivalent personal protection to all three LLINs. Relative to IRS plus untreated nets, community level protection is slightly higher when Olyset or PermaNet 2.0 nets are added onto IRS with pirimiphos methyl or lambda cyhalothrin but not DDT, and when Icon Life nets supplement any of the IRS insecticides. Adding IRS onto any net modestly enhances communal protection when pirimiphos methyl is sprayed, while spraying lambda cyhalothrin enhances protection for untreated nets but not LLINs. Addition of DDT reduces communal protection when added to LLINs. Where transmission is mediated primarily by An. arabiensis, adding IRS to high LLIN coverage provides only modest incremental benefit (e.g. when an organophosphate like pirimiphos methyl is used), but can be redundant (e.g. when a pyrethroid like lambda cyhalothin is used) or even regressive (e.g. when DDT is used for the IRS). Relative to IRS plus untreated nets, supplementing IRS with LLINs will only modestly improve community protection. Beyond the physical protection that intact nets provide, additional protection against transmission by An. arabiensis conferred by insecticides will be remarkably small, regardless of whether they are delivered as LLINs or IRS. The insecticidal action of LLINs and IRS probably already approaches their absolute limit of potential impact upon this persistent vector so personal protection of nets should be enhanced by improving the physical integrity and durability. Combining LLINs and non-pyrethroid IRS in residual transmission systems may nevertheless be justified as a means to manage insecticide resistance and prevent potential rebound of not only An. arabiensis, but also more potent, vulnerable and historically important species such as Anopheles gambiae and Anopheles funestus

Malaria elimination in Zanzibar: where next?

In 2018, Zanzibar developed a national malaria strategic plan IV (2018-2023) to guide elimination of malaria by 2023. We assessed progress in the implementation of malaria activities as part of the end-term review of the strategic plan. The review was done between August and October 2022 following the WHO guideline to assess progress made towards malaria elimination, effectiveness of the health systems in delivering malaria case management; and malaria financing. A desk review examined available malaria data, annual work plans and implementation reports for evidence of implemented malaria activities. This was complemented by field visits to selected health facilities and communities by external experts, and interviews with health management teams and inhabitants to authenticate desk review findings. A steady increase in the annual parasite incidence (API) was observed in Zanzibar, from 2.7 (2017) to 3.6 (2021) cases per 1,000 population with marked heterogeneity between areas. However, about 68% of the detected malaria cases were imported into Zanzibar. Malaria case follow-up and investigation increased from <70% in 2017 to 94% and 96% respectively, in 2021. The review noted a 3.7-fold increase of the health allocation in the country's budget, from 31.7 million USD (2017/18) to 117.3 million USD (2022/23) but malaria allocation remained low (<1%). The varying transmission levels in the islands suggest a need for strategic re-orientation of the elimination attempts from a national-wide to a sub-national agenda. We recommend increasing malaria allocation from the health budget to ensure sustainability of malaria elimination interventions

Long-Lasting Control of Anopheles arabiensis by a Single Spray Application of Micro-encapsulated Pirimiphos-methyl (Actellic(R) 300 CS).

Pyrethroid-resistant mosquitoes are an increasing threat to malaria vector control. The Global Plan for Insecticide Resistance Management (GPIRM) recommends rotation of non-pyrethroid insecticides for indoor residual spraying (IRS). The options from other classes are limited. The carbamate bendiocarb and the organophosphate pirimiphos-methyl (p-methyl) emulsifiable concentrate (EC) have a short residual duration of action, resulting in increased costs due to multiple spray cycles, and user fatigue. Encapsulation (CS) technology was used to extend the residual performance of p-methyl. Two novel p-methyl CS formulations were evaluated alongside the existing EC in laboratory bioassays and experimental hut trials in Tanzania between 2008-2010. Bioassays were carried out monthly on sprayed substrates of mud, concrete, plywood, and palm thatch to assess residual activity. Experimental huts were used to assess efficacy against wild free-flying Anopheles arabiensis, in terms of insecticide-induced mortality and blood-feeding inhibition. In laboratory bioassays of An. arabiensis and Culex quinquefasciatus both CS formulations produced high rates of mortality for significantly longer than the EC formulation on all substrates. On mud, the best performing CS killed >80% of An. arabiensis for five months and >50% for eight months, compared with one and two months, respectively, for the EC. In monthly bioassays of experimental hut walls the EC was ineffective shortly after spraying, while the best CS formulation killed more than 80% of An. arabiensis for five months on mud, and seven months on concrete. In experimental huts both CS and EC formulations killed high proportions of free-flying wild An. arabiensis for up to 12 months after spraying. There was no significant difference between treatments. All treatments provided considerable personal protection, with blood-feeding inhibition ranging from 9-49% over time. The long residual performance of p-methyl CS was consistent in bioassays and experimental huts. The CS outperformed the EC in laboratory and hut bioassays but the EC longevity in huts was unexpected. Long-lasting p-methyl CS formulations should be more effective than both p-methyl EC and bendiocarb considering a single spray could be sufficient for annual malaria control. IRS with p-methyl 300 CS is a timely addition to the limited portfolio of long-lasting residual insecticides