Cooperia pectinata (Nematoda: Trichostrongylidae) in New Zealand

Cooperia pectinata, a nematode parasite hitherto undetected in New Zealand, was recovered from the small intestine of a six-month-old red deer (Cervus elaphus) born on a deer farm in Taihape. The source of this infection is unknown but it seems likely that it originated from feral stock.     

Persistent efficacy of doramectin injectable against artificially induced infections with Cooperia punctata and Dictyocaulus viviparus in cattle.

Three studies were conducted to evaluate the persistent efficacy of doramectin injectable solution against experimental challenges with infective larvae of Cooperia punctata and Dictyocaulus viviparus. In each study, four groups of ten randomly-assigned calves, negative for trichostrongyle-type eggs on fecal examination, were treated subcutaneously in the midline of the neck with saline (1 ml/50 kg) on Day 0 or doramectin (200 microg/kg = 1 ml/50 kg) on Day 0, 7, or 14. Two additional calves from the same pool of animals were randomly assigned as larval-viability monitors and received no treatment. On Days 14-28, approximately 1000 and 50 infective larvae of Cooperia spp. and D. viviparus, respectively, were administered daily by gavage to each animal in Groups T1-T4. On Day 28, the two larval-viability monitor calves were inoculated in a similar manner with a single dose of approximately 30000 and 2000 larvae of Cooperia spp. and D. viviparus, respectively. Equal numbers of calves from each treatment group were killed on Days 42-45, as well as the two viability monitor animals to enumerate worm numbers. A 2% or 5% aliquot of small intestinal contents and washings were examined for worm quantification and identification, while 100% of the lung recoveries were quantified and identified. For each study and across the three studies, geometric mean worm recoveries for each treatment group were calculated from the natural log transformed data (worm count + 1) and were used to estimate percentage reduction. In the three studies, doramectin injectable solution was 97.5% efficacious against lungworms for up to 28 days and was 99.8% efficacious in reducing infection resulting from challenge with infective larvae of C. punctata for at least 28 days post-treatment.     

Corpus alienum in the abomasum of cattle.

In a field study consisting of about 7000 adult cattle 36 cases of corpora aliena (CAA) in the abomasum were found. After recording the clinical symptoms such as recurrent slight ruminal tympany and abdominal pain after eating, the metal specimens were found by a detector, with which the sound of the moving metal could be followed behind the xiphoid cartilage region. The diagnoses were confirmed by operation in 10 cases, when the specimens were found in the abomasum and removed. Four of these operations were performed directly near the linea alba and in six cases the specimens were removed by hand and magnet through the reticulo-abomasal opening after attempting the routine hard ware operation. In an additional 16 cases the foreign body could not be caught hold of and removed by a ruminal operation due to passage difficulties through the reticulo-abomasal opening. The results of the direct or indirect operations when the CAA was removed, were good. It is concluded that CAA can occur in some 5 per mille of the patient population and in some 50 per cent of these cases a traumatic reticulitis is also found. In most cases the clinical symptoms are vague. Only if the symptoms are painful and recurrent is the operation indicated.     

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-12p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC-1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (L3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P<0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P<0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-12p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by T(H)2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group.     

Assessment of biosecurity level in small-scale broiler farms in the Western highlands of Cameroon (Central Africa)

The recent outbreak of avian influenza in the poultry sector of Cameroon has raised a concern about the level of implementation of biosecurity measures (BM) on poultry farms. Therefore, a study using a questionnaire on 102 randomly selected poultry farms was undertaken. Some measures with high adoption levels (>?90%) included “protection of airing openings of poultry barns by a wire mesh,” “no external animal allowed to enter the farm,” “respect of all-in all-out principle,” “functional footbath,” “rodent control,” and “feedstuff protection from access to rodents.” The least implemented measures (less than 20%) were “wearing of dedicated clothing (clean coveralls and boots) by workers,” “disinfection of visitors,” and “presence of storage room for dead animals.” Only for “isolation” component of biosecurity, compliance with biosecurity measures was good, with more than 50% of farms having an adoption rate greater than 75%. The mean biosecurity score for the assessed farms was 19.29?±?1.89 for a maximum score of 38 points. The score was significantly and positively correlated (p?<?0.05) with the number of chicken and the geographic location of farms. The other results showed that the mean number of broiler per cycle was 550.82?±?76 for a stocking density of 27.20?±?6.64 birds per m². In conclusion, broiler production in the Western highland is dominated by small-scale units with low level of biosecurity practices positively and significantly (p?<?0.05) influenced by some production characteristics. To improve biosecurity practices in the area, efforts must focus on measures concerning “sanitation” and “traffic control” components of biosecurity.     

Risk factors of Salmonella infection in laying hens in Menoua Division,Western region of Cameroon (Central Africa)

Salmonella infections in poultry farms are overlooked in many African countries; yet these infections are mostly zoonotic with impact on both poultry industry and public health. Considering the impact of Salmonella in laying hens, and the role of laying hens as a source of Salmonella outbreak in human, knowledge of the status of Salmonella on laying hen farms as well as the factors influencing the presence of Salmonella is important. In a cross sectional study, cloacal swabs were collected from 270 commercial laying hens on 27 farms located in Menoua Division. These samples were cultured on standard media. A questionnaire was used to collect information on animals, farms and farmer’s characteristics. The prevalence of Salmonella was 93.34%; three zoonotic isolates namely S. Enteritidis (75.90%), S. Paratyphi (11.90%), and S. Typhimurium (5.60%) were identified. The location of farms was significantly associated with presence of Salmonella, and the risk of infection was 10-fold higher in Nkong-ni than Santchou (p < 0.05). Other potential risk factors such as flock size, age of the farm (infrastructure), or water source were not associated with Salmonella infection. The prophylactic measures against avian diseases in the country must include measures against Salmonella to protect poultry industry and public health.     

Ineffikacy of ivermectin against Cooperia spp. infection in cattle

Ivermectin, either as an injectable or topical formulation, is used widely to control parasites in New Zealand cattle. These formulations have been well documented as having high efficacy against the major nematode species which infect tattle^(1)(2)(3)(4), although the efficacy against adult Cooperia oncophora is reported to be 95%⁽⁵⁾. This is considered to be one of the doselimiting species. There have been two reports of lower than expected efficacy of the topical formulation against Cooperia spp. in cattle. In a slaughter trial, Bisset et al. ⁽⁶⁾ found that ivermectin applied topically at the recommended dose rate of 500 𝛍g/kg failed to achieve a statistically significant reduction in the burden of Cooperia spp. worms compared with untreated calves. The geometric mean of adult Cooperia spp. worms present in the treated calves was 5788 compared with 7524 present in the untreated calves. This reduced efficacy was not attributed to acquired anthelmintic resistance because ivermectin had not been used on the farm previously. In Australia, Eagleson and Allertonc⁽⁷⁾ found in controlled field trials that ivermectin applied topically at 500 𝛍g/kg ranged in efficiency in reducing faecal egg counts from 80.7% to 100% ; four out of 13 trials achieving less than 90% and a further four were in the range of 90–95% reduction. Larvae obtained from the faeces of ivermectin-treated cattle on days 11–14 after treatment were predominantly Cooperia spp.     

Abundance and survival of infective larvae of the cattle nematodes Cooperia punctata, Haemonchus placei and Oesophagostomum radiatum from faecal pats in a wet tropical climate

Observations were made on the abundance and survival of Haemonchus placei, Cooperia punctata and Oesophagostomum radiatum infective larvae from cattle faecal pats exposed at various times of the year in north Queensland wet tropics. Pats exposed in the hot, wet season yielded abundant larvae on herbage. In the dry season, although low numbers of infective larvae were usual, considerable numbers were produced under conditions of heavy dews on dense herbage. Irrespective of season of deposition of pats, the resulting larvae persisted generally for not longer than 10 to 12 weeks, and in large numbers for only 2 to 6 weeks. The findings suggest that prevention of contamination in the wet season, and in the dry season when light rainfalls are accompanied by heavy dews on dense herbage, will result in low levels of larval infestation on herbage. Rotational grazing in the area is suggested as a means of worm control.     

Ultrastructural study of globidian parasites infecting the abomasum of sheep in Germany.

Globidian parasites infecting the abomasum of sheep in Germany were investigated by means of electron microscopy. The frequency of infection was found to be 93 %. The globidian cyst-like bodies contained multinucleate schizonts, developing merozoites or fully developed merozoites. Among the latter there were two different types, namely short and long forms. The process of merozoite formation was described in detail. The giant schizonts were subdivided into multinucleate cell portions of irregular size and shape. Their nuclei were then arranged at the periphery of the cell portions and underwent their last division which was combined with the differentiation of merozoites. The long form merozoites were elongated cylindrical in shape with terminal nucleus. They measured 7.7 μm in length and 1.0 μm in width. The merozoites of the short type were spindle-shaped with a central nucleus. They were 5.0 μm long and 1.0 μm wide. The globidian parasites were located in a parasitophorous vacuole of an intact host cell.     

A sero-epidemiological survey of blood parasites in cattle in the north-eastern Free State, South Africa

A survey to determine the incidence of parasites in cattle (n = 386) was conducted in the north eastern Free State between August 1999 and July 2000. Giemsa-stained blood smears were negative for blood parasites. A total of 94% of the cattle were sero-positive for Babesia bigemina by indirect fluorescent antibody test while 87% were sero-positive for Anaplasma by enzyme-linked immunosorbent assay. The observation of negative blood smears but high incidence of positive serological results for Anaplasma and Babesia for the same group of cattle indicates that this area is endemic for these diseases but with a stable disease situation. All the animals were sero-negative for B. bovis and this is probably because the tick vector (Boophilus microplus) which transmits the disease is not present in the Free State Province. Two tick species belonging to the family ixodidae were found on cattle, namely Boophilus decoloratus and Rhipicephalus evertsi evertsi. In the present study significant differences in seasonal burdens of B. decoloratus occurred, with the highest infestations recorded from February to June. The presence of R. evertsi evertsi throughout the year without any or with small fluctuations in winter months was observed, with a peak from February to May.     

Dynamics of the humoral immune response of calves infected and re-infected with Cooperia punctata

The dynamics of the humoral immune response of calves were analysed after primary infection and re-infection with the intestinal nematode Cooperia punctata. 12 male 5 month-old Holstein-Friesian calves were randomly divided into two groups A and B. At the beginning of the experiment Group A animals were each infected experimentally with a single oral dose of 130,000 infective third stage larvae (L3) of C. punctata. The animals of Group B were kept as non-infected controls. The two calves from Group A with the highest infections died of cooperiosis at 32 and 44 days after infection (DAI), respectively. On DAI 100 the calves were treated with the recommended dose of oxfendazole. On DAI 180 the remaining four calves of Group A and three animals of Group B (B1) were infected with 260,000 L3 of C. punctata, while the other three calves of Group B (B2) served as non-infected controls. Monitoring of the humoral immune response predominantly demonstrated an IgG1 response against both adult and L3 antigen of C. punctata. Moreover, re-infections increased the levels of these immunoglobulins. IgA levels were less increased than IgG1 and no significant increase was observed in IgG2 and IgM levels. Immunoblotting analysis showed that total IgG present in the serum of the primary infected animals mainly reacted against adult proteins of 12-14 and 17-20 kDa and against L3 proteins of 33 and 43 kDa. After re-infection total IgG reacted with the same adult proteins but also with an adult 29 kDa protein.     

Benzimidazole-resistant beta-tubulin alleles in a population of parasitic nematodes (Cooperia oncophora) of cattle

Three anthelmintic classes with distinct mechanisms of action are commercially available. Selection of nematode populations resistant to all these drugs has occurred, particularly in trichostrongyloid parasites of sheep. Anthelmintic resistance in cattle parasites has only recently been recognized and appears to be less pronounced, even though very similar species infect both hosts. To understand the bases for differences in the rate of resistance development in sheep versus cattle parasites, it is important to first demonstrate that the same kinds of resistance alleles exist in both. The benzimidazoles (BZ), which have been used for more than 40 years, were chosen as an example. BZ-sensitive (BZ(S)) and BZ-resistant (BZ(R)) nematodes that parasitize sheep have been distinguished at the molecular level by a single nucleotide change in the codon for amino acid 200 of a beta-tubulin gene, a switch from TTC (phenylalanine) to TAC (tyrosine). PCR primers were designed to completely conserved regions of trichostrongyloid beta-tubulin genes and were used to amplify DNA fragments from Haemonchus contortus (cDNA from a BZ(S) and a BZ(R) library) as positive controls. The technique was then extended to the cattle parasites, Cooperia oncophora and Ostertagia ostertagi (from genomic DNA). Sequence analysis proved the presence of amplified BZ(S) alleles in all three species and BZ(R) alleles in the BZ(R) population of H. contortus. Based on these data, nested PCR primers using the diagnostic T or A as the most 3' nucleotide were designed for each species. Conditions for selective PCR were determined. To demonstrate feasibility, genomic DNA was recovered from individual H. contortus L(3) larvae from both BZ(S) and BZ(R) populations. Genomic DNA was also isolated from >70 individual adult male C. oncophora collected from a cattle farm in New Zealand with reported BZ resistance. Allele-specific PCR discriminated among heterozygotes and homozygotes in both species. This method could find utility in studying the molecular epidemiology of BZ resistance in cattle parasites and for defining the variables that limit the development and spread of anthelmintic resistance in this host.