In an attempt to establish an ideal method for mass production of Calicophoron microbothrium metacercariae, a study was carried out to compare the shedding capacities of Bulinus tropicus naturally and experimentally infected with C. microbothrium. A total of 906 F1 B. tropicus between 4 and 5 weeks old were each experimentally infected with two C. microbothrium miracidia and monitored for 12 weeks. The infected snails were fed on dried lettuce and fish flakes and were kept in 1 l plastic aquaria housed in a snail room where temperature, light and humidity were controlled. Seventy-four percent of the experimentally infected snails died during the prepatent period and of the remaining, only 13.2 % developed patent infection, while 12.5 % were refractory. Snail growth rate was poor and the average shedding rate was 20 cercariae per snail per day. Compared to the experimentally infected snails, 2 200 adult B. tropicus, collected from the field and naturally infected with C. microbothrium, yielded high numbers of metacercariae. Eighty-four percent of the snails died within 7 weeks of the study with peak mortality occurring from the 2nd to the 4th week of infection and coinciding with an overall decrease in the number of cercariae shed.

In an attempt to establish an ideal method for mass production of Calicophoron microbothrium metacercariae, a study was carried out to compare the shedding capacities of Bulinus tropicus naturally and experimentally infected with C. microbothrium. A total of 906 F1 B. tropicus between 4 and 5 weeks old were each experimentally infected with two C. microbothrium miracidia and monitored for 12 weeks. The infected snails were fed on dried lettuce and fish flakes and were kept in 1 l plastic aquaria housed in a snail room where temperature, light and humidity were controlled. Seventy-four percent of the experimentally infected snails died during the prepatent period and of the remaining, only 13.2 % developed patent infection, while 12.5 % were refractory. Snail growth rate was poor and the average shedding rate was 20 cercariae per snail per day. Compared to the experimentally infected snails, 2 200 adult B. tropicus, collected from the field and naturally infected with C. microbothrium, yielded high numbers of metacercariae. Eighty-four percent of the snails died within 7 weeks of the study with peak mortality occurring from the 2nd to the 4th week of infection and coinciding with an overall decrease in the number of cercariae shed.

During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000) in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16 264 (calves 5 418; weaners 5 461 and adults 5 385) faecal samples were collected during the entire period of the study and 2 500 (15.4 %) of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P 0.001), for adult cattle than calves ( P 0.01) and in the wet season over the dry season (P 0.01). Faecal egg output peaked from August / September to March / April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P 0.01). The snail population was low between December and March and started to increase in April reaching a peak in September / October. The number of juvenile snails peaked between April and August. The mean number of snails collected was negatively correlated with rainfall and positively correlated with temperature. Mean number of snails collected was also positively correlated with Potamogeton plant species and negatively correlated with Cyperus plant species. However, none of the L. natalensis collected from the habitats were found shedding Fasciola cercariae. Metacercariae were found on herbage from the fringes of the snail habitats between February and August for both years, with most of the metacercariae concentrated on herbage 0-1 m from the banks of the habitats. Based on the findings of this study, anthelmintic treatment should be administered in December / January to control chronic and mature fasciolosis. A second treatment should be given in April / May to reduce pasture contamination and subsequently snail infection, as this is the time the snail population starts to build up. To control acute fasciolosis due to the immature liver flukes a third treatment should be given in August. The first application of molluscicides to control the snail intermediate hosts can be done in June the time when the snail is harbouring the parasite and a second application in September in order to kill new generations of infected snails.

During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000) in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16 264 (calves 5 418; weaners 5 461 and adults 5 385) faecal samples were collected during the entire period of the study and 2 500 (15.4 %) of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001), for adult cattle than calves ( P < 0.01) and in the wet season over the dry season (P < 0.01). Faecal egg output peaked from August / September to March / April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P < 0.01). The snail population was low between December and March and started to increase in April reaching a peak in September / October. The number of juvenile snails peaked between April and August. The mean number of snails collected was negatively correlated with rainfall and positively correlated with temperature. Mean number of snails collected was also positively correlated with Potamogeton plant species and negatively correlated with Cyperus plant species. However, none of the L. natalensis collected from the habitats were found shedding Fasciola cercariae. Metacercariae were found on herbage from the fringes of the snail habitats between February and August for both years, with most of the metacercariae concentrated on herbage 0-1 m from the banks of the habitats. Based on the findings of this study, anthelmintic treatment should be administered in December / January to control chronic and mature fasciolosis. A second treatment should be given in April / May to reduce pasture contamination and subsequently snail infection, as this is the time the snail population starts to build up. To control acute fasciolosis due to the immature liver flukes a third treatment should be given in August. The first application of molluscicides to control the snail intermediate hosts can be done in June the time when the snail is harbouring the parasite and a second application in September in order to kill new generations of infected snails.

The present study investigated the distribution of nerves in the oviduct of the sexually immature ostrich. The presence of protein gene product 9.5, neurofilament protein and neuron specific enolase nerve fibres were demonstrated in the infundibulum, magnum, isthmus, shell gland and vagina. Nerve fibres containing protein gene product 9.5, neuron specific enolase and neurofilament protein were particularly numerous in the tunica muscularis and intermuscular connective tissue areas of the shell gland and vagina. The presence of a large number of nerves in these oviductal regions is probably important in the coordination of muscle contraction. An interesting finding of the study was the presence of protein gene product 9.5 and neuron specific enolase immunopositive nerve fibres in the walls of blood vessels. It is likely that these nerves are autonomicin nature and play a role in the regulation of blood flow. This study has shown the presence of an extensive neural network in the oviduct of the ostrich. In addition, the results of the investigation have shown that the neuronal markers protein gene product 9.5, neurofilament protein and neuron specific enolase can be used to demonstate nerve fibres in the ostrich.

The present study investigated the distribution of nerves in the oviduct of the sexually immature ostrich. The presence of protein gene product 9.5, neurofilament protein and neuron specific enolase nerve fibres were demonstrated in the infundibulum, magnum, isthmus, shell gland and vagina. Nerve fibres containing protein gene product 9.5, neuron specific enolase and neurofilament protein were particularly numerous in the tunica muscularis and intermuscular connective tissue areas of the shell gland and vagina. The presence of a large number of nerves in these oviductal regions is probably important in the coordination of muscle contraction. An interesting finding of the study was the presence of protein gene product 9.5 and neuron specific enolase immunopositive nerve fibres in the walls of blood vessels. It is likely that these nerves are autonomicin nature and play a role in the regulation of blood flow. This study has shown the presence of an extensive neural network in the oviduct of the ostrich. In addition, the results of the investigation have shown that the neuronal markers protein gene product 9.5, neurofilament protein and neuron specific enolase can be used to demonstate nerve fibres in the ostrich.

For nearly 50 years the ixodid tick Hyalomma marginatum turanicum, reputedly introduced into South Africa on imported Persian sheep, has been considered identical to the Asian Hyalomma (Euhyalomma) marginatum turanicum Pomerantzev, 1946. Comparisons of this tick with the Asian H. (E.) m. turanicum and other subspecies of Hyalomma (Euhyalomma) marginatum, however, reveal that it is an old taxon, namely Hyalomma rufipes glabrum Delpy, 1949. It is hereby reinstated as Hyalomma (Euhyalomma) glabrum, and its adults are redescribed and its immature stages described for the first time. The preferred hosts of its adults are large herbivores such as zebras, gems bok and eland, on which it occurs during summer. The preferred hosts of its immature stages are scrub hares and ground-frequenting birds, on which it is present during autumn and winter. Data on its distribution and possible disease relationships are also provided.

For nearly 50 years the ixodid tick Hyalomma marginatum turanicum, reputedly introduced into South Africa on imported Persian sheep, has been considered identical to the Asian Hyalomma (Euhyalomma) marginatum turanicum Pomerantzev, 1946. Comparisons of this tick with the Asian H. (E.) m. turanicum and other subspecies of Hyalomma (Euhyalomma) marginatum, however, reveal that it is an old taxon, namely Hyalomma rufipes glabrum Delpy, 1949. It is hereby reinstated as Hyalomma (Euhyalomma) glabrum, and its adults are redescribed and its immature stages described for the first time. The preferred hosts of its adults are large herbivores such as zebras, gems bok and eland, on which it occurs during summer. The preferred hosts of its immature stages are scrub hares and ground-frequenting birds, on which it is present during autumn and winter. Data on its distribution and possible disease relationships are also provided.

The ultrastructure of the follicular wall in primordial, previtellogenic and vitellogenic follicles of the sexually immature ostrich is described in the present study. The follicular wall consists of a zona radiata, granulosa cell layer, basal lamina and thecal layer. Cytoplasmic processes from the plasma membranes of the granulosa cell layer and the ovocyte form the zona radiata in previtellogenic and vitellogenic follicles. The granulosa cell layer transforms from simple cuboidal epithelium in primordial follicles to simple columnar or pseudostratified columnar epithelium in previtellogenic and vitellogenic follicles. Transosomes were observed along the apical and lateral plasma membranes of granulosa cells. The thecal layer in previtellogenic and vitellogenic follicles consists of interna and externa components. The fibroblasts in the theca externa contain microfilaments, which are thought to be actin filaments. The study revealed ultrastructural features, which are associated with the transportation of yolk precursors and nutrients into the ovoplasm. In addition, the study indicates that, although the cells in the theca externa contain microfilaments, they do not possess the ultrastructural characteristics of smooth muscle cells.

The ultrastructure of the follicular wall in primordial, previtellogenic and vitellogenic follicles of the sexually immature ostrich is described in the present study. The follicular wall consists of a zona radiata, granulosa cell layer, basal lamina and thecal layer. Cytoplasmic processes from the plasma membranes of the granulosa cell layer and the ovocyte form the zona radiata in previtellogenic and vitellogenic follicles. The granulosa cell layer transforms from simple cuboidal epithelium in primordial follicles to simple columnar or pseudostratified columnar epithelium in previtellogenic and vitellogenic follicles. Transosomes were observed along the apical and lateral plasma membranes of granulosa cells. The thecal layer in previtellogenic and vitellogenic follicles consists of interna and externa components. The fibroblasts in the theca externa contain microfilaments, which are thought to be actin filaments. The study revealed ultrastructural features, which are associated with the transportation of yolk precursors and nutrients into the ovoplasm. In addition, the study indicates that, although the cells in the theca externa contain microfilaments, they do not possess the ultrastructural characteristics of smooth muscle cells.

Oestrous synchronization involves synchronization of ovarian follicular turnover, new wave emergence, and finally induction of ovulation. The final step can be synchronized by the parenteral administration of either GnRH or oestradiol benzoate. This study investigated corpus luteum and follicular emergence after ovulation had been induced by the administration of either GnRH or oestradiol benzoate. The injection of oestradiol benzoate may have delayed the emergence of the first follicular wave subsequent to the induced ovulation; administration of oestradiol benzoate or GnRH lowered the progesterone rise so that the maximum dioestrous concentration of progesterone on Day 9 was lower when cows were treated during pro-oestrus compared to the spontaneously ovulating controls. One implication of findings from the present study is that induction of ovulation with either oestradiol benzoate or GnRH, administered 24 or 36 h after withdrawal of the CIDR device, respectively, may lower fertility. Future studies must identify the timing of administration relative to the time of CIDR device withdrawal and the optimum concentration of oestradiol benzoate or GnRH that would not have untoward effects on the development of the corpus lutea, particularly within the first week of dioestrus.

Oestrous synchronization involves synchronization of ovarian follicular turnover, new wave emergence, and finally induction of ovulation. The final step can be synchronized by the parenteral administration of either GnRH or oestradiol benzoate. This study investigated corpus luteum and follicular emergence after ovulation had been induced by the administration of either GnRH or oestradiol benzoate. The injection of oestradiol benzoate may have delayed the emergence of the first follicular wave subsequent to the induced ovulation; administration of oestradiol benzoate or GnRH lowered the progesterone rise so that the maximum dioestrous concentration of progesterone on Day 9 was lower when cows were treated during pro-oestrus compared to the spontaneously ovulating controls. One implication of findings from the present study is that induction of ovulation with either oestradiol benzoate or GnRH, administered 24 or 36 h after withdrawal of the CIDR device, respectively, may lower fertility. Future studies must identify the timing of administration relative to the time of CIDR device withdrawal and the optimum concentration of oestradiol benzoate or GnRH that would not have untoward effects on the development of the corpus lutea, particularly within the first week of dioestrus.

A serological survey was conducted in apparently healthy, unvaccinated indigenous Tswana goats and sheep in Kasane, Maun and Shakawe districts in northwestern Botswana in order to determine in these animals, the levels of exposure to the South African Territories (SAT) serotypes: SAT 1, SAT 2 and SAT 3 of foot-and-mouth disease virus (FMDV). A total of 250, 142 and 134 goat sera originating respectively from Kasane, Maun and Shakawe districts were tested for FMDV antibodies against the three SAT serotypes by the liquid phase blocking enzyme-linked immunosorbent assay and 26 of 250 (10.4 %), 5 of 142 (3.5 %) and 18 of 134 (13.4 %) were positive either to SAT 1 or SAT 3, or to both serotypes. None of the goats' sera was positive to SAT 2 serotype. All sheep sera (n = 9) tested negative against all three serotypes of the virus. The findings are discussed in relation to results of other serological surveys carried out elsewhere.

A serological survey was conducted in apparently healthy, unvaccinated indigenous Tswana goats and sheep in Kasane, Maun and Shakawe districts in northwestern Botswana in order to determine in these animals, the levels of exposure to the South African Territories (SAT) serotypes: SAT 1, SAT 2 and SAT 3 of foot-and-mouth disease virus (FMDV). A total of 250, 142 and 134 goat sera originating respectively from Kasane, Maun and Shakawe districts were tested for FMDV antibodies against the three SAT serotypes by the liquid phase blocking enzyme-linked immunosorbent assay and 26 of 250 (10.4 %), 5 of 142 (3.5 %) and 18 of 134 (13.4 %) were positive either to SAT 1 or SAT 3, or to both serotypes. None of the goats' sera was positive to SAT 2 serotype. All sheep sera (n = 9) tested negative against all three serotypes of the virus. The findings are discussed in relation to results of other serological surveys carried out elsewhere.

The tortoise tick Amblyomma marmoreum was collected from large numbers of reptiles and other animals during the course of numerous surveys conducted in South Africa. A total of 1 229 ticks, of which 550 were adults, were recovered from 309 reptiles belonging to 13 species, with leopard tortoises, Geochelone pardalis being the most heavily infested. The 269 birds sampled harboured 4 901 larvae, 217 nymphs and no adult ticks, and the prevalence of infestation was greatest on hel meted guinea fowls, Numida meleagris. Only two larvae were recovered from 610 rodents, including 31 spring hares, Pedetes capensis, whereas 1 144 other small mammals yielded 1 835 immature ticks, of which 1 655 were collected from 623 scrub hares, Lepus saxatilis. The 213 carnivores examined harboured 2 459 ticks of which none were adult. A single adult tick and 6 684 larvae and 62 nymphs were recovered from 656 large herbivores, and a total of 4 081 immature ticks and three adults were collected from 1 543 domestic animals and 194 humans. Adult male and female A. marmoreum were most numerous on reptiles during January and February, and larvae during March. The largest numbers of larvae were present on domestic cattle and helmeted guineafowls in the Eastern Cape Province during March or April respectively, whereas larvae were most numerous on helmeted guineafowls, scrub hares and the vegetation in north-eastern Mpumalanga Province during May. In both provinces nymphs were most numerous between October and December. Amblyomma marmoreum appears to be most prevalent in the western regions of the Western and Eastern Cape and Free State provinces, and the north-eastern regions of the Northern Cape, KwaZulu- Natal, Mpumulanga and Limpopo provinces.

The tortoise tick Amblyomma marmoreum was collected from large numbers of reptiles and other animals during the course of numerous surveys conducted in South Africa. A total of 1 229 ticks, of which 550 were adults, were recovered from 309 reptiles belonging to 13 species, with leopard tortoises, Geochelone pardalis being the most heavily infested. The 269 birds sampled harboured 4 901 larvae, 217 nymphs and no adult ticks, and the prevalence of infestation was greatest on hel meted guinea fowls, Numida meleagris. Only two larvae were recovered from 610 rodents, including 31 spring hares, Pedetes capensis, whereas 1 144 other small mammals yielded 1 835 immature ticks, of which 1 655 were collected from 623 scrub hares, Lepus saxatilis. The 213 carnivores examined harboured 2 459 ticks of which none were adult. A single adult tick and 6 684 larvae and 62 nymphs were recovered from 656 large herbivores, and a total of 4 081 immature ticks and three adults were collected from 1 543 domestic animals and 194 humans. Adult male and female A. marmoreum were most numerous on reptiles during January and February, and larvae during March. The largest numbers of larvae were present on domestic cattle and helmeted guineafowls in the Eastern Cape Province during March or April respectively, whereas larvae were most numerous on helmeted guineafowls, scrub hares and the vegetation in north-eastern Mpumalanga Province during May. In both provinces nymphs were most numerous between October and December. Amblyomma marmoreum appears to be most prevalent in the western regions of the Western and Eastern Cape and Free State provinces, and the north-eastern regions of the Northern Cape, KwaZulu- Natal, Mpumulanga and Limpopo provinces.

Rapid Rural Appraisal methods were used to collate and code the indigenous knowledge on animal healthcare of Tsonga speaking people of South Africa. There was a rapport between local disease names as described by their clinical signs by the farmers and the local veterinary services important disease list. The perceived causes of diseases were physico-biological elements and no reference to ancestral guidance was recorded. Males and old people were more knowledgeable but females and young people did show a certain degree of confidence during general discussions. Plants were more frequently used than other non-conventional remedies with cattle being the most treated animals. Farmers reported using 19 plant species belonging to 12 families. Plants were collected from the wild when needed and no specific storage system was used. They were administered as decoctions or infusions of single plants. These remedies were used not only as alternatives to expensive pharmaceutical products but also because in certain diseases or chronic cases, they were thought to be more efficacious.

Rapid Rural Appraisal methods were used to collate and code the indigenous knowledge on animal healthcare of Tsonga speaking people of South Africa. There was a rapport between local disease names as described by their clinical signs by the farmers and the local veterinary services important disease list. The perceived causes of diseases were physico-biological elements and no reference to ancestral guidance was recorded. Males and old people were more knowledgeable but females and young people did show a certain degree of confidence during general discussions. Plants were more frequently used than other non-conventional remedies with cattle being the most treated animals. Farmers reported using 19 plant species belonging to 12 families. Plants were collected from the wild when needed and no specific storage system was used. They were administered as decoctions or infusions of single plants. These remedies were used not only as alternatives to expensive pharmaceutical products but also because in certain diseases or chronic cases, they were thought to be more efficacious.

Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59 % and 67 %, respectively. In a second experiment, a stabilate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35 % per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35 % loss of infectivity.

Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59 % and 67 %, respectively. In a second experiment, a stabilate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35 % per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35 % loss of infectivity.