Distribution and histopathological changes induced by cysts of Taenia solium in the brain of pigs from Tanzania.

Neurocysticercosis (NCC) caused by Taenia solium cysts is a frequent but neglected parasitic disease of the central nervous system (CNS) worldwide. The aim of this study was to describe anatomical locations of cysts in the CNS and the corresponding inflammation. A total of 17 naturally infected pigs were used to evaluate the distribution of cysts and, of these, seven were used to evaluate the corresponding inflammation further, through histopathology. Clinical signs in the pigs included dullness, sluggishness, somnolence, apathy and loss of consciousness. Cysts were distributed in all cerebral lobes, i.e. 39.7% in the frontal lobe, 20.3% in the parietal lobe, 20.0% in the occipital lobe and 19.7% in the temporal lobe, and only 0.4% in the cerebellum. No cysts were found in the spinal cord. Cysts were localized as follows: 47.9% in the dorsal subarachnoid, 46.9% in the parenchyma, 4.4% in the subarachnoid base and 0.9% in the ventricles. The results of the histopathology revealed lesions in an early inflammatory stage, i.e. stage I, in all anatomical locations except for two, which showed more of an inflammatory reaction, stage III, in one pig. It was concluded that clinical signs in pigs were neither pathognomonic nor consistent. These signs, therefore, cannot be used as a reliable indicator of porcine NCC. Furthermore, T. solium cysts were found to be in abundance in all cerebral lobes, and only a few were found in the cerebellum. Regarding the inflammatory response, no significant differences were found in the location and total number of cysts. Thus, further studies are needed to explain the determinants of cyst distribution in the CNS and assess in detail clinical signs associated with porcine NCC.

Distribution and histopathological changes induced by cysts of Taenia solium in the brain of pigs from Tanzania.

Neurocysticercosis (NCC) caused by Taenia solium cysts is a frequent but neglected parasitic disease of the central nervous system (CNS) worldwide. The aim of this study was to describe anatomical locations of cysts in the CNS and the corresponding inflammation. A total of 17 naturally infected pigs were used to evaluate the distribution of cysts and, of these, seven were used to evaluate the corresponding inflammation further, through histopathology. Clinical signs in the pigs included dullness, sluggishness, somnolence, apathy and loss of consciousness. Cysts were distributed in all cerebral lobes, i.e. 39.7% in the frontal lobe, 20.3% in the parietal lobe, 20.0% in the occipital lobe and 19.7% in the temporal lobe, and only 0.4% in the cerebellum. No cysts were found in the spinal cord. Cysts were localized as follows: 47.9% in the dorsal subarachnoid, 46.9% in the parenchyma, 4.4% in the subarachnoid base and 0.9% in the ventricles. The results of the histopathology revealed lesions in an early inflammatory stage, i.e. stage I, in all anatomical locations except for two, which showed more of an inflammatory reaction, stage III, in one pig. It was concluded that clinical signs in pigs were neither pathognomonic nor consistent. These signs, therefore, cannot be used as a reliable indicator of porcine NCC. Furthermore, T. solium cysts were found to be in abundance in all cerebral lobes, and only a few were found in the cerebellum. Regarding the inflammatory response, no significant differences were found in the location and total number of cysts. Thus, further studies are needed to explain the determinants of cyst distribution in the CNS and assess in detail clinical signs associated with porcine NCC.

Helminth parasites of the Kafue lechwe antelope ( Kobus leche kafuensis): a potential source of infection to domestic animals in the Kafue wetlands of Zambia.

The Kafue lechwe antelope (Kobus leche kafuensis), a medium-sized, semi-aquatic antelope, grazes extensively on pastures accessed by livestock in and around Lochinvar and Blue Lagoon national parks in the Kafue wetlands of Zambia. This interaction has a potential for bi-modal transmission of a wide range of parasitic helminths between lechwe and domestic ruminants. A survey was conducted to investigate the status of helminths in the Kafue lechwe during the 2008 (July-December) hunting season, involving 65 animals hunted under special research licences. Worm identification was based on morphological features using standard identification keys. Eleven different types of helminths were identified in the animals studied; namely, Oesophagostomum, Bunostomum, Cooperia, Dictyocaulus, Marshallagia, Stilesia, Setaria, Trichuris, Fasciola, amphistomes and Schistosoma. Amphistomes (100%) and Oesophagostomum (60.9%) were the most common while Fasciola (7.8%) and Stilesia (1.6%) were the least of the identified helminths. There was no evidence that helminths, at intensities observed, adversely affected the health of the lechwe. The degree of worm infection was observed to vary between the two study areas, with Blue Lagoon recording higher infection levels compared to Lochinvar. The host range of many of the helminths found in the Kafue lechwe is broad and could serve as a potentially stable source of infection to domestic animals such as goats and cattle. Therefore, issues concerning livestock management and conservation may arise.

The immune response in Taenia solium neurocysticercosis in pigs is associated with astrogliosis, axonal degeneration and altered blood-brain barrier permeability.

Immunohistochemistry was used to examine the immuno-pathological changes and the extent of neuronal damage caused by either viable or dead Taenia solium cysticerci during porcine neurocysticercosis. Thirty pig brains with cerebral cysticercosis and 5 brains from T. solium free pigs were used in this study. Results revealed extensive astrogliosis, neuronal and mostly axonal damage in both early (grade I) and late (grades III and V) lesions as evidenced by an increased expression of glial fibrillary acidic protein (GFAP) and neurofilament protein (NFP). In many late lesions, astrocyte end-feet formed glial scars that surrounded the dead parasite. Rapid angiogenesis resulted in blood vessels lacking astrocyte end-feet suggesting loss of blood-brain barrier (BBB) hence allowing an influx of peripheral blood immune cells such as eosinophils, macrophages, CD3+ T cells, B lymphocytes and plasma cells into lesions. This study showed that porcine NCC was associated with severe nervous tissue damage, the host response of which is a collaborative effort between the local and peripheral immune responses comparable to that observed in human NCC. Results further implied that porcine NCC could be a useful model for understanding the course of NCC in human as well as provide useful information for therapeutic and/or immune strategies.

Host-cell apoptosis in Taenia solium-induced brain granulomas in naturally infected pigs.

To assess whether apoptosis occurs in pig brain granulomas due to Taenia solium cysticerci, brain tissues from 30 pigs naturally infected with T. solium cysticercosis were evaluated by terminal deoxynucleotidyl transferase-end labelling (TUNEL) staining. In addition, tissues were stained with CD3 marker to identify T lymphocytes. Examination of TUNEL-stained tissues showed apoptotic cells in early lesions that contained viable cysticerci. Apoptotic cells were primarily found interspersed with normal cell types, and were mostly located in the inflammatory infiltrate. Late or advanced granulomas with disintegrated scolices did not show TUNEL-positive cells. CD3+ cells were found in both early and advanced lesions and apoptosis mainly co-localized with CD3+ T lymphocytes. This suggests that these cells are constantly undergoing apoptosis and thus die as soon as they arrive at the site of infection. Apoptosis indeed may be one way by which T. solium cysticerci down-regulate the host's cellular immune response in early cysticercosis. Therefore, further research is needed to establish if other cells besides T-lymphocytes are also a target for destruction by cysticerci in early cysticercosis as well as studies to assess if cysteine protease is expressed by viable cysticerci in situ.

Taenia solium porcine cysticercosis: viability of cysticerci and persistency of antibodies and cysticercal antigens after treatment with oxfendazole.

The aim of this study was to assess the effect of treating Taenia solium infected pigs with oxfendazole (OFZ) on viability and clearance of cysticerci and the corresponding persistence of specific antibody isotypes (IgG(total), IgG1, IgG2 and IgA) and circulating cysticercal antigen (CCA). Antibody isotypes and CCA responses were measured by antibody-ELISA (Ab-ELISA) and antigen ELISA (Ag-ELISA), respectively. Correlations were made between antibodies, CCA and the total number of cysticerci enumerated at necropsy. Forty pigs with cysticercosis were randomly allocated into two groups: Treatment group (n=20) was treated with OFZ at 30 mg/kg orally while the treatment control group (n=20) was not treated. Five uninfected pigs served as negative controls. Pigs were killed at 1, 4, 8 and 26 weeks post-treatment (wkpt). Overall, the mean total cyst count in treated pigs was 2904+/-5397 (mean+/-S.D.) while in the controls it was 6235+/-6705. Mean cyst viability was 5+/-11% (mean+/-S.D.) and 97+/-4% in treated and control pigs, respectively. Results showed that OFZ killed muscular cysticerci over a period of 4 weeks but failed to kill cerebral cysticerci. Antibodies, CCA responses and clearance of dead cysts from the meat, depended on the cyst intensity of individual pigs at time of treatment since both antibody and CCA correlated with intensity of cysticerci at necropsy (r=0.441, P=0.005; r=0.654, P<0.001), respectively. IgG1 responses were the best indicator of treatment efficacy because they were predominant in both infected treated and control pigs and disappeared early after treatment. Both Ab/Ag-ELISA failed to detect cysts in the brain. Though dead cysticerci took some time (26 wkpt) to clear from the meat, treatment of porcine cysticercosis with OFZ should, in combination with other intervention measures be considered as an important, cost-effective measure in the control of taeniosis/cysticercosis.

Risk factors associated with porcine cysticercosis in selected districts of Eastern and Southern provinces of Zambia.

To determine the risk factors associated with Taenia solium transmission in humans and pigs in the rural areas of Eastern and Southern provinces of Zambia, a questionnaire was administered in 788 households from 155 villages. Pigs were examined from 800 households. Tongue examination and enzyme-linked immunosorbent assay (Ag-ELISA) for the detection of circulating antigens of T. solium cysticerci were used to measure infection in pigs. A snowballing technique was utilised to select households with pigs. Prevalence of households with pigs infected with T. solium on tongue examination by district ranged from 12.7% to 32.1% with Ag-ELISA having a range of 30.0-51.7%. Of the total number of households visited, 18.8% and 37.6% had at least one pig positive for porcine cysticercosis on tongue examination and Ag-ELISA, respectively. Risk factors associated with T. solium infection were lack of pork inspection at slaughter (96.7%), consumption of pork with cysts (20.1%), selling of pork infected with T. solium cysticerci (18.3%), free-range husbandry system (83.2%) and absence of latrines (58.0). Free-range husbandry system (OR=1.68; 95% CI=1.36-2.07) was a significant risk factor for porcine cysticercosis in the surveyed areas. The result that pigs were mostly kept on free-range and semi-intensive husbandry systems may have permitted them to have access to eating human faeces that could be contaminated with tapeworm eggs. This study has shown that T. solium infection poses a high public health risk in the study areas and urban areas as well. We recommend that a human survey be conducted to verify the human exposure to taeniasis and/or cysticercosis in Zambia.

Control of Taenia solium taeniasis/cysticercosis: The best way forward for sub-Saharan Africa?

Taenia solium taeniasis/cysticercosis is a neglected parasitic zoonosis with significant economic and public health impacts. Control measures can be broadly grouped into community health education, improvements in hygiene and sanitary conditions, proper meat handling at household and community level, improved standards of meat inspection, pig management, treatment of individual patients and possibly human populations, and treatment and/or vaccination of porcine populations. This manuscript looks critically into currently existing control options and provides suggestions on which (combination of) tools would be most effective in the control of T. solium taeniasis/cysticercosis in sub-Saharan Africa. Field data and disease transmission simulations suggest that implementation of a single intervention control strategy will not lead to a satisfactory reduction of disease morbidity or transmission. A feasible strategy to combat T. solium taeniasis/cysticercosis would include a combination of approaches focussing on both human (health education and treatment) and animal host (management, treatment and vaccination), which can vary for different communities and different geographical locations. Selection of the specific strategy depends on cost-effectiveness analyses based on solid field data, currently unavailable, though urgently needed; as well as on health priorities and resources of the country. A One Health approach involving medical, veterinary, environmental and social sectors is essential for T. solium to be controlled and eventually eliminated. Finally the success of any intervention is largely dependent on the level of societal and political acceptance, commitment and engagement.

Comparative fluke burden and pathology in condemned and non-condemned cattle livers from selected abattoirs in Zambia.

After dissecting 70 condemned and 32 non-condemned cattle livers collected from Lusaka, Chisamba, Mongu and Senanga abattoirs and Turnpike slaughter slab, significantly higher numbers of liver flukes (Fasciola gigantica) (P < 0.001) were found in the condemned livers (mean +/- SD = 100.6 +/- 16.7) than in the non-condemned livers (mean +/- SD = 0.7 +/- 0.5). Liver flukes found in 9.4% of the non-condemned livers suggest that abattoir records of liver inspection may underestimate F. gigantica infections. Average faecal fluke egg counts from animals with condemned livers (5 eggs per gram [EPG]) were significantly higher (P < 0.001) than in animals with non-condemned livers (0.8 EPG). No correlation was found between egg counts and number of flukes. Fibrosis and calcification were common in condemned livers, being severest in the vicinity of the bile ducts. Only two (6.3%) of the non-condemned livers showed pathological changes on the liver edges. The severe liver damage and high worm burden may explain low production levels experienced in cattle in Zambia maintained under traditional systems of management where worm control and good management programmes are rarely practiced.

Seasonal pattern of bovine fasciolosis in the Kafue and Zambezi catchment areas of Zambia.

In order to establish seasonality of bovine fasciolosis in the Southern province of Zambia, 288 cattle were examined at slaughter at Turnpike slaughter slab for 1 year. The examination involved liver inspection and coprological examination. Liver condemnation rates were relatively high at the beginning of both the cold dry season and the rainy season (May/June and December, respectively). All cattle, regardless of age, had higher fluke abundances in the post-rainy season (39.1% young and 42.1% adult) while the lowest rates were in the hot dry season (13.3% young and 14.3% adult). On coprological examination, the highest abundance was in the post-rainy season (45.0%) and the lowest in the cold dry season (24.9%). From November (end of dry season) to February/March (end of rains), more fluke eggs were found than in any other period. The distribution of fluke eggs was significantly different (p<0.001) among the four seasons. Identified associations were: egg counts were highest in post-rainy (mean abundance=146) and lowest in cold dry season (mean abundance=118) at 95% confidence (CI 0.64-1.25). Differences in abundance observed according to origin, sex and age of cattle were not significant on both methods. Based on these results, it may be concluded that Fasciola gigantica was present throughout the year in cattle in Zambia but the abundance was highest in the post-rainy season and lowest in the cold dry season.

Taenia solium metacestode preparation in rural areas of sub-Saharan Africa: a source for diagnosis and research on cysticercosis.

BACKGROUND: Taenia solium metacestodes/cysts obtained from pig carcasses constitute a primary source for diagnostic tools used for the detection of human cysticercosis. Data on T. solium cyst preparation in Africa is still scarce but required to establish independent reference laboratories. OBJECTIVES: The aim of the present study is a) to present the likely yield of T. solium cyst material by the use of two different preparation methods in the field and b) to investigate its suitability for immunodiagnosis of human cysticercosis. METHODS: In Zambia, Uganda and Tanzania 670 pigs were screened for T. solium infection. Cysts were prepared by 'shaking method' and 'washing method'. Generated crude antigens were applied in a standard western blot assay. RESULTS: 46 out of 670 pigs (6.9%) were found positive for T. solium (Zambia: 12/367, 3.3%; Uganda: 11/217, 5.1%; Tanzania 23/86, 26.7%). Mean values of 77.7 ml whole cysts, 61.8 ml scolices/membranes and 10.9 ml cyst fluid were obtained per pig. Suitability of collected material for the use as crude antigen and molecular diagnostic techniques was demonstrated. CONCLUSION: This study clearly shows that T. solium cyst preparation in African settings by simple field methods constitutes an effective way to obtain high quality material as source for diagnostic tools and research purposes.

Short Communication: In Vitro Efficacy Testing of Praziquantel, Ivermectin, and Oxfendazole against Taenia Solium Cysts.

Oxfendazole is recommended as the drug of choice for treating porcine cysticercosis. The drug does not kill brain cysts and is not registered for use in pigs. Latest its safety in the recommended dose has been questioned. The aim of this study was to investigate two alternative anthelminthics. The efficacy of praziquantel and ivermectin was compared to oxfendazole In Vitro on Taenia solium. Cysts of T. solium were isolated from infected pork and incubated in culture media together with the drugs. The degree of evagination was used as effect measurement and determined after 6 hours. Praziquantel had a half maximal effective concentration (EC(50)) of value 0.006 ± 0.001 µg/mL. Ivermectin did not show any impact on the evagination in concentrations from 0.001 to 30 µg/mL and neither did oxfendazole in concentrations from 0.001 to 50 µg/mL.

Porcine cysticercosis in southeast Uganda: seroprevalence in kamuli and kaliro districts.

The recent recognition of neurocysticercosis as a major cause of epilepsy in Uganda and changes in pig demography have lead to a need to better understand the basic epidemiology of Taenia solium infections in pigs and humans. Human exposure is a function of the size of the animal reservoir of this zoonosis. This is the first field survey for porcine cysticercosis to investigate the prevalence of antigen-positive pigs across an entire rural district of south-east Uganda. In our field surveys, 8.6% of 480 pigs screened were seropositive for the parasite by B158/B60 Ag-ELISA. In addition, of the 528 homesteads surveyed 138 (26%) did not have pit latrines indicating a high probability of pigs having access to human faeces and thus T. solium eggs. This study thus indicates the need for better data on this neglected zoonotic disease in Uganda, with a particular emphasis on the risk factors for infection in both pigs and humans. In this regard, further surveys of pigs, seroprevalence surveys in humans and an understanding of cysticercosis-related epilepsy are required, together with risk-factor studies for human and porcine infections.

Prevalence of fasciolosis in Zambian cattle observed at selected abattoirs with emphasis on age, sex and origin.

The aim of this study was to determine the prevalence and some factors influencing occurrence of fasciolosis in cattle, carried out at selected major abattoirs in Zambia. Of 841 cattle livers inspected and 677 faecal samples analysed, prevalence rates of 53.9% and 48.9%, respectively, were found. Cumulative prevalence of 60.9% (n = 677) was recorded. According to age, no significant difference of infection was found. Female cattle on liver inspection (59.3%) and coprological examination (65.2%) had significantly higher (P < 0.001) rates than males (44.5% and 36.3% respectively). Origin of cattle had a significant influence (P < 0.001) on the prevalence rate. According to fluke egg count classification, 68.5% of cattle had light infection, 20.9% moderate, 4.4% heavy and 6.2% severe. These results indicate that Fasciola gigantica infection is an important condition that leads to high liver condemnations and/or trimmings in cattle tendered for slaughter.