Babesiosis is a tick borne disease (TBD) caused by parasites of the genus Babesia, with 13 considerable worldwide economic, medical, and veterinary impact. Babesiosis and other 14 TBDs were considered responsible for 50% of the deaths of cattle occurred in 15Mozambique in the first year after importation from neighbouring countries. 16Here, we present the detection of B. bigemina and B. bovis in cattle from Mozambique 17 and the development of a novel PCR method. For this study, blood samples were 18 collected in one farm located near Maputo city. The samples were analyzed using a 19 previously described nested PCR and a novel hot-start PCR method. Primers were 20 selected for the hot-start PCR based on the putative gene of an undescribed aspartic 21 protease named babesipsin, present in both B. bovis and B. bigemina. The combination of 22 hot start polymerase and long primers (29-31bp) were in this study determinant for the 23 Manuscript 2 successful amplification and detection in only one PCR. The babesipsin hot-start PCR is 24 a fast, simple, and sensitive method, with some advantages over current methods. A total 25of 117 field samples were tested by hot-start PCR, and 90 were positive for B. bigemina 26 (77%), 82 were positive for B. bovis (70%), 61 were mixed infections (52%) and only 6 27 were negative (5.1%). The results confirm that this area of Mozambique is endemic for 28 Babesiosis, and that this TBD should be regarded as a threat for imported cattle. 29 30

Molecular detection of Babesia species in apparently healthy cattle within an endemic region was carried out in order to determine the prevalence of carriers and the geographical distribution of Babesia bigemina and Babesia bovis in Maputo Province, Mozambique. Samples from 477 animals at 5 localities were analysed using 2 techniques, the semi-nested hot-start PCR and the reverse line blot (RLB) assay. With the semi-nested hot-start PCR, detection of B. bigemina ranged between 30% and 89%, and of B. bovis between 27% and 83%. The RLB assay was comparatively less sensitive in this study and detection of B. bovis ranged from 0% to 17%, and B. bigemina was not detected at all by this technique. Analysis of new sequences of the 18S rRNA gene revealed that the current B. bigemina RLB probe is not specific for the identification of isolates in Mozambique. The RLB assay, however, resulted in the detection of 8 other haemoparasite species belonging to the genera Babesia, Theileria, Anaplasma and Ehrlichia. 18S rRNA gene sequences from the Theileria spp. were identified, and a phylogenic tree constructed with these sequences yielded a heterogeneous T. mutans-like group. In conclusion, infection with B. bigemina and B. bovis is endemic in Maputo Province, but rates of transmission vary. Furthermore, mixed infections with the haemoparasites responsible for several tick-borne diseases in cattle are common in Mozambique.

Background . Microbiological quality of drinking water supplied in Moamba, a small town in southern Mozambique, was assessed by collecting and analyzing 91 water sample from 5 sampling sites: raw or inlet water, treated water and 3 household taps along the water distribution system. The presence of Escherichia coli as indicator fecal contamination, three bacterial pathogens, Vibrio cholerae, Salmonella and Campylobacter spp., and Cefotaximee resistant E. coli as antibiotic resistance determinant, was assessed. Results . The results showed fecal contamination in all types of water samples: E. coli was found in 100% of inlet water samples, in 21% of treated water samples, and in 22% of tap water samples. No Salmonella spp. was detected during the study. The presence of V. cholerae was detected in 42% of all water samples tested: 100% of inlet water samples, in 16% of treated water samples, and in 23% household tap water samples. All V. cholerae confirmed isolates where genotyped by PCR as non-O1/non-O139; however, 9 isolates showed the presence of the genes encoding for cholera toxin. The presence of Campylobacter spp. was detected in 36% of the water samples tested: in 95% of inlet water samples, in 10% of treated water samples and in 23% household tap water samples. Cefotaxime resistant E. coli was detected in 63% of inlet water, 16% of treated water, and in 9% of tap water samples, these isolates were also resistant to multiple other antibiotics: ampicillin, streptomycin, tetracycline chloramphenicol. All 70 V. cholerae non-O1/non-O139 confirmed isolated were resistant to ampicillin, 51% to streptomycin, 13% to gentamycin, and 1 isolate was resistant to tetracycline; 13% showed a multi-drug resistant profile, being resistant to at least three antibiotics. Conclusion . The presence of fecal contamination and pathogens in the water treatment system and household taps in Moamba indicates a health risk for the population. This burden increases by the presence of bacterial pathogens showing multidrug resistance.

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