Control Strategies for Carcinogenic-Associated Helminthiases: An Integrated Overview.

Helminthiases are extremely prevalent in the developing world. In addition, the chronic infection with some parasitic worms are classified as carcinogenic. Therefore, it is utmost importance to understand the parasite-host interactions, the mechanisms underlay carcinogenesis and how they could be counteracted. This knowledge may ultimately guide novel control strategies that include chemotherapy-based approaches targeting these pathogens and associated pathologies caused by their infections. Little is known on how some helminthiases are associated with cancer; however, it has been hypothesized that chemical carcinogenesis may be involved in the process. Here, we summarize the current knowledge on chemical carcinogenesis associated with helminthiases, along with available therapeutic options and potential therapeutic alternatives including chemotherapy and/or immunotherapy. Ideally, the treatment of the carcinogenic helminthiases should target both the parasite and associated pathologies. The success of any chemotherapeutic regimen often depends on the host immune response during the infection and nutritional status among other factors. The close association between chemotherapy and cell-mediated immunity suggests that a dual therapeutic approach would be advantageous. In addition, there is a pressing need for complementary drugs that antagonize the carcinogenesis process associated with the helminth infections.

rTgOWP1-f, a specific biomarker for Toxoplasma gondii oocysts.

Toxoplasma gondii oocyst wall protein 1 (TgOWP1) integrates a family of seven proteins, consensually assumed as specific antigens of Toxoplasma gondii oocyst stage, located in the outer layer of the oocyst wall. Herein, we notice the expression of a recombinant antigen, rTgOWP1-f, derived from a fragment selected on basis of its structural homology with Plasmodium MSP1-19. Rabbit polyclonal antibodies anti-rTgOWP1-f evidence ability for specific identification of environmental T. gondii oocysts. We assume, rTgOWP1-f, as a possible biomarker of oocysts. In addition, we present findings supporting this vision, including the development of an immunodetection method for T. gondii oocysts identification.

Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits.

BACKGROUND: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts. METHODS: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light. RESULTS: Forty percent of the analysed samples (95% CI: 25.5-56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6-179.9 oocysts/g. CONCLUSIONS: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts.

Esophageal cancer in Mozambique: should mycotoxins be a concern?

Fumonisin B1 (FB1) is a mycotoxin frequently found in agricultural commodities. The toxin poses a considerable risk for human and animal health. FB1 is among several mycotoxins produced by Fusarium spp. contaminating virtually any cereal and other Poaceae. Their intracellular action includes the promotion of oxidative stress through the generation of reactive oxygen species (ROS) that damage biomolecules such as DNA. These toxic effects were observed in vivo and in vitro. However, the association between esophageal lesions and oxidative stress induced by FB1. Studies in China, Iran and South Africa showed higher exposure to fumonisins in areas with higher risk of esophageal cancer (EC). Exposure to mycotoxins may be inevitable in Mozambique. How mycotoxins, particularly fumonisins from the contaminated food, can be associated with the emergence of EC in Mozambique? Herein, we revise the literature and present some pieces of evidence in order to highlight the burden of mycotoxins and to provide evidence-based considerations for the stakeholders involved in the management of the EC agenda in Mozambique. The information presented herein supports the need to implement novel and/or to revisit the existent detoxification methods to reduce the global burden of mycotoxins and its outcomes in health management.

Potent cationic antimicrobial peptides against Mycobacterium tuberculosis in vitro.

BACKGROUND: Tuberculosis (TB) is known to be one of the 10 causes of global death by infectious agents. The increasing numbers of multiple antibiotic resistance (MDR-TB) and cases of extensive resistance to antibiotics (XDR-TB) have led to the development of new and effective TB therapy. Cationic antimicrobial peptides (CAMPs) have emerged in the research as a safe and effective treatment against a variable range of bacterial and fungi pathogens, including Mycobacterium tuberculosis (M. tuberculosis). METHOD: This study developed a new CAMP coupled with cinnamic acid derivatives, and studied the antimicrobial activity against clinical isolates of M. tuberculosis (H37Rv) and MDR-TB. RESULTS: All modified CAMPs showed enhanced activity against both M. tuberculosis strains and were capable of disrupting heavy clumping of mycobacteria in culture. In addition, all modified CAMPs were able to substantially inhibit the intracellular growth of both strains at low concentrations. CONCLUSIONS: The characteristic proprieties of cinnamic acid+CAMP(n) successfully inhibited the growth of both clinical isolates M. tuberculosis and MDR-TB in vitro and have, for now, promising use as a drug adjuvant due to their effect on mycobacteria growth.

Estrogen-like metabolites and DNA-adducts in urogenital schistosomiasis-associated bladder cancer.

An estrogen-DNA adduct mediated pathway may be involved in the pathogenesis of the squamous cell carcinoma of the bladder associated with infection with the blood fluke Schistosoma haematobium. Extracts from developmental stages of S. haematobium, including eggs, induce tumor-like phenotypes in cultured cells. In addition, estrogen-derived, reactive metabolites occur in this pathogen and in sera of infected persons. Liquid chromatography-mass spectrometry analysis was performed on urine from 40 Angolans diagnosed with urogenital schistosomiasis (UGS), half of who also presented UGS-associated squamous cell carcinoma and/or urothelial cell carcinoma. The analysis revealed numerous estrogen-like metabolites, including seven specifically identified in UGS cases, but not reported in the database of metabolites in urine of healthy humans. These schistosome infection-associated metabolites included catechol estrogen quinones (CEQ) and CEQ-DNA-adducts, two of which had been identified previously in S. haematobium. In addition, novel metabolites derived directly from 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG) were identified in urine of all 40 cases of UGS. These metabolites can be expected to provide deeper insights into the carcinogenesis UGS-induced bladder cancer, and as biomarkers for diagnosis and/or prognosis of this neglected tropical disease-linked cancer.

Clinically relevant multidrug resistant Salmonella enterica in swine and meat handlers at the abattoir.

The presence of multidrug resistant (MDR) Salmonella serotypes in slaughtered swine, carcasses, meat and meat handlers is scarcely evaluated. Recently we demonstrated that diverse Salmonella serotypes are frequently present in swine, pork meat and carcasses, and meat handlers at Portuguese abattoirs. Here we have characterized their antibiotic resistance phenotypes and genotypes, helping elucidate the flow of MDR Salmonella in the food chain. Testing 60 Salmonella isolates from different serotypes, the highest frequencies of resistance were observed for tetracycline (T) [70% (n=42/60), tet(A)/tet(B)/tet(G)], streptomycin (S) [63% (n=38/60), aadA2/strA/strB], sulfamethoxazole (Sul) [62% (n=37/60), sul1/sul2/sul3] and ampicillin (A) [57% (n=34/60), blaPSE-1/blaTEM]. Thirty-seven percent (n=22/60) carried class 1 integrons and multidrug resistance was frequently observed (63% n=38/60), including those serotypes common to human infections [S. Typhimurium 78% n=25/32; S. 4,[5],12:i:- 67% n=2/3; S. Rissen 75% (n=3/4); S. London 67% n=2/3; S. Derby 55%; n=6/11)]. The emergent S. 4,[5],12:i:- isolates were mostly characterized by ASSuT phenotype [blaTEM/strA-strB/sul2/tet(B)], typical of the European clone, while for the first time the ST phenotype [strA-strB-tet(A)-tet(B)] was also observed. Moreover, we report a first finding of a MDR phenotype in S. London [ANSSuT; blaTEM-strA-strB-sul2-tet(A)]. Our findings suggest that the abattoir environment and the slaughter operations seem not only to harbor MDR serotypes that originated in the pig reservoir, but also propagate them through cross-contamination processes, involving meat handlers. The present study suggests a probable relationship between swine and human salmonellosis throughout the food chain, which is of interest for epidemiological, animal health and public health purposes.

Schistosoma haematobium and bladder cancer: what lies beneath?

Schistosoma haematobium is a parasitic flatworm that infects millions of people, mostly in the developing world, and is associated with high incidence of bladder cancer although why is not clear. But our group was able to define the mechanistic relationship for the first time between infection of S. haematobium and cancer. We used in vitro models to demonstrate the presence of informative carcinogenesis-associated phenotypes in CHO cells exposed to Sh total antigen, in which we showed increased cell proliferation, decreased apoptosis, up regulation of the anti-apoptotic molecule Bcl-2, down regulation of the tumor suppressor protein p27, and increased cell migration and invasion. We further discuss the molecular and cellular events that might be responsible for schistosomiasis-related bladder cancer.

Tumourigenic effect of Schistosoma haematobium total antigen in mammalian cells.

Schistosoma haematobium is endemic in several regions of Africa and has been shown to be associated with predominantly squamous cell bladder carcinoma. The mechanisms underlying the association between S. haematobium and bladder squamous cell carcinoma is largely unknown. All the reports so far, demonstrate exclusively an epidemiological evidence linking S. haematobium infection with squamous cell bladder carcinoma. We hypothesized that these parasite antigens might induce tumourigenesis. For this, we used normal mammalian cells of Chinese hamster ovary (CHO) and treated the cells in culture with S. haematobium total antigen (Sh). Our results showed increased proliferation in Sh-treated cells in comparison with the controls. The CHO cells exposed to Sh were inoculated subcutaneously into male nude mice and formed sarcomas (n = 5/5). The cells from the sarcomas expressed vimentin filaments and were negative to cytokeratin. Our results demonstrate for the first time that S. haematobium antigens induce tumour development in nude mice.

Detection of Cryptosporidium spp. and Giardia duodenalis in surface water: a health risk for humans and animals.

The objective of the present study was to determine the degree of contamination by Cryptosporidium spp. and Giardia duodenalis in a river basin in a livestock farming area in Galicia (NW, Spain). Water samples (50 l) were collected at 22 points in the main basin (including 5 recreational areas), and at the source and mouth of the 3 most important rivers and at the mouth of a smaller, secondary river. Faecal samples were collected from dairy cattle selected at random from 18 herds farmed in the area. A total of 139 neonatal calves, 480 heifers and 697 cows were sampled. The prevalence, intensity of infection and the risk associated with the spread of infection by both enteropathogens were determined. Water and faecal samples were collected in spring, summer, autumn and winter of 2007. The species and genotypes of these parasites present in the water samples were identified. In both water and faecal samples, more parasitic stages were collected in spring and summer than in autumn and winter. In spring, Cryptosporidium spp. oocysts were detected in 33 (9.4%) cows from 13 (72.2%) herds, and G. duodenalis cysts were detected in 56 (16.0%) cows from 15 farms (83.3%); the intensity of infection ranged from 5 to 7895 G. duodenalis cysts per gram of faeces. Infective stages of Cryptosporidium spp. and G. duodenalis were also detected in respectively 26 (89.6%) and 27 (93.1%) water samples, in spring. The mean concentrations of parasites ranged from 2 to 1200 Cryptosporidium spp. oocysts per litre and from 2 to 400 G. duodenalis cysts per litre. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. The presence of both protozoans must be monitored in cattle, in sources of water used for recreational purposes and in artificial waterways used by farmers (water channels, animal drinking water and drainage systems).

Schistosoma haematobium total antigen induces increased proliferation, migration and invasion, and decreases apoptosis of normal epithelial cells.

Schistosome worms are blood-dwelling flukes that cause chronic infection in more than 200 million people and are thought to be responsible for 500,000 deaths annually. During infection with Schistosoma haematobium, eggs are deposited in the mucosa and submucosa of the bladder and lower ureters. Squamous cell carcinoma (SCC) of the bladder is a long-term sequela of chronic infection. The mechanisms underlying the association between S. haematobium and SCC of the bladder are largely unknown, with all reports to date exclusively demonstrating epidemiological evidence linking S. haematobium infection with SCC of the bladder. We hypothesised that the parasite antigens might induce alterations in epithelial cells towards cancer. For this we used Chinese Hamster Ovary (CHO) cells and treated the cells in culture with S. haematobium total antigen (Sh). Our results showed increased proliferation, increased S-phase and decreased apoptosis, as well as down-regulation of tumour suppressor p27 and up-regulation of anti-apoptotic molecule Bcl-2 in Sh-treated cells compared with controls. We also found increased migration and invasion. To our knowledge, this is the first report demonstrating alterations of normal epithelial cells as a direct effect of S. haematobium antigens.

Molecular characterization of Cryptosporidium and Giardia isolates from cattle from Portugal.

Fecal samples from 291 calves and 176 adult cattle in Northern Portugal were screened for Cryptosporidium and Giardia using a formalin-ethyl acetate concentration method. Acid-fast staining techniques for Cryptosporidium oocyst identification and direct microscopic observation of fecal smears for Giardia cyst identification were performed so as immunofluorescence microscopy examination. Polymerase chain reaction methods were employed to determine the genotype of each isolate. Molecular characterization was performed using amplification and sequencing of the hsp70 and 18SrRNA genes of Cryptosporidium and beta-giardin gene and glutamate dehydrogenase for assemblage determination of Giardia duodenalis. Seventy-four out of 291 calves (25.4%) and 8 out of 176 adult bovines (4.5%) were positive for Cryptosporidium. Forty-one out of 291 calf samples (14.1%) and 1 out of 176 adults samples (0.57%) were positive for Giardia. From the Cryptosporidium positive samples we obtained 63 isolates from calves samples and 7 isolates from adult samples. Additionally, Giardia was isolated in 13 out of 41 positive samples from calves and it was also possible to isolate Giardia from the positive adult sample. Molecular characterization of the Cryptosporidium and Giardia isolates showed us that C. parvum and G. duodenalis assemblage E were the prevalent species. C. parvum may infect humans, representing a potential public health risk. On the other hand, the assemblages B and A2 of Giardia, previously described in humans, were here identified in cattle. Further studies will be needed for determine the importance of cattle as carrier of zoonotic assemblages of G. duodenalis.