Protection against high-dose homologous infection in calves immunized with intestine or membrane extracts from Haemonchus placei.

Control of Haemonchus placei, one of the most important cattle nematodes in Brazil, relies on the use of anthelmintics. However, there is a need for integrated control, which includes active immunization. The aim of this work was to assess the protection afforded to calves by immunization with adult H. placei extracts against a high-dose challenge infection, a condition frequently found in the tropics. Holstein calves aged 8-10 months were immunized four times with intestinal extracts (Group D) or with a Triton X-100-soluble fraction of adult H. placei (Group A), challenge-infected with 120,000 infective larvae and sacrificed 40 days later. Immunized animals had higher IgG titers than the controls against tested fractions after the 2nd immunization, peaking after the 4th. Sera from both immunized groups recognized bands of similar apparent mass in both antigenic preparations, some of which were similar in molecular weight to Haemonchus contortus antigens with known protective effect to sheep. Egg counts were 49% and 57% lower in Groups A and D than in controls, respectively. High levels of protection were observed in two of the four calves in Group D, as evidenced by very low worm numbers recovered at necropsy, absence of eggs in the uteri of the recovered females and reduced worm length. Group D animals also showed milder signs of anemia than the other infected animals. Results demonstrate that protection against homologous high-dose challenge can be achieved by immunizing calves with H. placei gut antigens.

A family of activation associated secreted protein (ASP) homologues of Cooperia punctata.

Activation-associated secreted proteins (ASP) of nematodes have been studied as potential vaccine components. In this study we report the cloning and analysis of cDNA and genomic sequences of Cooperia punctata and establish the presence of two 75% identical ASP-1 genes in C. punctata. Additional C. punctata ASP paralogues were shown to be present. Analysis of PCR products amplified from genomic DNA from a pool of worms revealed extensive sequence diversity within this family of proteins, reflecting the presence of different ASP paralogues in a single worm as well as extensive polymorphisms between different worms. ASP proteins contain a conserved region called the sperm-coating protein (SCP) domain of unknown function, which is present as a single copy in proteins from yeast and a wide range of multi-cellular organisms. Only in three nematodes has a protein composed of duplicated SCP-domains been identified. C. punctata is the first organism in which at least two such genes are found. Database searches identified similarity of the C-terminal cysteine-rich domain of ASP proteins to a nematode metallothionein motif. Cp-asp-1b was expressed in Escherichia coli and both the N-terminal and C-terminal domain were shown to be recognized by sera of C. punctata infected bovines. The description of the asp gene family of C. punctata provides the basis for more detailed studies into the extent of variation and immunological recognition of this family that may assist in rational vaccine design.

Teaching of undergraduate veterinary parasitology: the South American perspective.

Undergraduate teaching of veterinary parasitology in South American countries is basically similar to most of the veterinary schools and offers lectures and practicals in parasitology (P) predominantly in the second year of the curriculum, and parasitic diseases (PD) in the third year, but in some schools also in the fourth and fifth years. However, there is a great variation in teaching hours allocated to P/PD between totals of about 50-169 h during the full curriculum. In addition to the lectures and practicals, schools are increasingly using tools (CD-ROMs) for self-instruction and new forms of education, such as workshops, round tables, and field practicals. The large number of important parasite species of large and small animals in tropical and subtropical areas would require additional hours for teaching--at least in some of the schools--considering the multidisciplinary characteristics of veterinary parasitology.

Cooperia punctata trickle infections: parasitological parameters and evaluation of a Cooperia recombinant 14.2 kDa protein ELISA for estimating cumulative exposure of calves.

Three groups of four calves each were trickle infected with three different levels of Cooperia punctata: 310 (group A), 1250 (group B) and 5000 (group C) third stage infective larvae (L3) twice a week over a 17-week period. Group D was the non-infected control group. Parasitological parameters as faecal egg counts (epg), worm burdens, size of worms and number of eggs per female were collected and the differences between the groups compared. Serological analyses were also conducted to investigate the efficiency of a recombinant C. oncophora CoES 14.2kDa protein in an ELISA to detect C. punctata antibodies. Group C had higher faecal egg counts until week 9 when the values decreased to those in group B. Mean faecal egg counts in group A were always lower than in the two other infected groups. The worm burdens were highest in group C, and lowest in group A, although the number of worms as a percentage of total larval intake was higher for the lower group. The mean length of the worms was shorter and the number of eggs per female lower for group C than for both other groups. ELISA using the CoES 14.2kDa proved to be efficient in measuring C. punctata antibodies. For group C it took 4 weeks to get increased levels of antibodies and this was one and 2 months more for groups B and A, respectively. Overall, there was a congruent relation between C. punctata antibodies and the cumulative exposure to the three different levels of trickle infections.

In vitro interaction of Brazilian strains of the Nematode-trapping fungi Arthrobotrys spp. on Panagrellus sp. and Cooperia punctata

In vitro tests were carried out to verify the activity of 26 Brazilian isolates of predatory fungi of the genus Arthrobotrys on a free-living nematode (Panagrellus sp.) and on infective larvae of Cooperia punctata, a parasitic gastrointestinal nematode of cattle. The results showed that the free-living nematode Panagrellus sp. was the most preyed upon, compared to C. punctata, for all the fungal treatments. Also, variable predatory capacity was observed for different fungal isolates belonging to the same genus when applied to different nematode species

In vitro activity of Brazilian strains of the predatory fungi Arthrobotrys spp. on free-living nematodes and infective larvae of Haemonchus placei

In vitro tests were carried out to assess the activity of 26 Brazilian isolates of predatory fungi of the genus Arthrobotrys on a free-living nematode (Panagrellus sp.) and on infective larvae of Haemonchus placei, a parasitic gastrointestinal nematode of cattle. The results showed that the free-living nematode Panagrellus sp. was the most preyed upon, compared to H. placei, for all the fungal treatments. Also, variable predatory capacity was observed for different fungal isolates belonging to the same genus when applied to different nematode species

Comparaçäo entre patogenia e susceptibilidade de bezerros Nelore e Holandês à infecçäo experimental com Haemonchus placei (Place, 1893) / Comparative susceptibility and pathogeny of Nelore and Holstein-Friesian calves to the experimental infection of Haemonchus placei (Place, 1893)

Dois grupos de cinco bezerros, das raças Holandesa e Nelore, com idade aproximada de nove meses, receberam por via oral uma dose única de 1.000 larvas infectantes (L3) por kg de peso corporal de Haemonchus placei. Os animais foram mantidos sob observaçäo clínica após a infecçäo. Amostras de sangue foram coletadas semanalmente por punçäo venosa, desde uma semana antes, até a oitava semana pós-infecçäo. Os estudos hematológicos abrangeram hematócrito, contagem diferencial de leucócitos, determinaçöes de hemoglobina, fibrinogênio e proteínas plasmáticas. Os exames parasitológicos compreenderam exames de fezes semanais com contagem de ovos (OPG) e estimativa de cargas de vermes na necropsia. Amostras de tecido da mucosa do abomaso foram submetidos a estudos histopatológicos. Ambos os grupos mostraram números crescentes de OPG após a quinta semana, sendo que os bezerros de raça Holandesa apresentaram maiores contagens do que os da raça Nelore. Na terceira semana pós-infecçäo e até o final do experimento, os bezerros da raça Holandesa tiveram anemia e hipoproteinemia, enquanto que os bezerros Nelore näo apresentaram alteraçöes significativas desses parâmetros. A necropsia, os bezerros da raça Holandesa apresentaram uma carga de vermes significativamente maior dos que os da raça Nelore. As alteraçöes patológicas visíveis e lesöes histopatológicas foram similares, embora macroscopicamente fossem levemente mais evidentes na raça Holandesa. Estes resultados demonstram que bezerros da raça Holandesa säo mais susceptíveis à infecçäo e aos efeitos patogênicos de H. placei do que bezerros da raça Nelore