Benzimidazole resistance in Nematodirus spathiger and N. filicollis in New Zealand

AIM: To determine the prevalence of benzimidazole resistance in Nematodirus spathiger and N. filicollis from a sample of New Zealand farms.

METHODS: The efficacy of albendazole (ABZ) against Nematodirus spp. was assessed by faecal nematode egg count reduction (FECR) tests undertaken in lambs aged 3–8 months old on 27 sheep farms throughout New Zealand. On each farm, groups of 10–16 lambs were either treated with ABZ (4.75?mg/kg) or remained as untreated controls. Faecal samples were collected from all animals at the time of treatment and 7–10 days later. Faecal nematode egg counts (FEC) were performed using a modified McMaster technique. Larvae were cultured from pooled faecal samples, collected 7–10 days after treatment from each group, by incubation at 20°C for 6 weeks, 4°C for 26 weeks then 13°C for 2 weeks. The resulting third stage larvae were identified to species using a multiplex PCR assay, that identified species-specific sequences in the second internal transcribed spacer region of ribosomal DNA. The efficacy of ABZ for N. spathiger and N. filicollis was calculated from the proportion of the two species in culture and the group mean FEC before and after treatment. Only farms with a mean of 10?epg for each species in untreated samples were included for analysis. Resistance was defined as an efficacy <95%.

RESULTS: On farms that met the threshold of 10?epg in faecal samples, benzimidazole resistance was found on 20/21 (95%) farms for N. spathiger compared with 4/10 (40%) farms for N. filicollis (p<0.05). In samples collected following treatment, a mean of 83 (min 46, max 100)% of Nematodirus spp. larvae recovered from the untreated groups were N. spathiger, compared with 94 (min 45, max 100)% in the ABZ treated groups (p=0.03). This change in percentage was not influenced by the overall efficacy of treatment based on the FECR test (p=0.324).

CONCLUSION: The results confirm the high level of resistance in N. spathiger in New Zealand and that benzimidazole resistance was more common in N. spathiger than N. filicollis. While resistance to benzimidazole anthelmintics has been reported previously in New Zealand, this is the first report of N. filicollis being resistant to benzimidazole anthelmintics.     

An evaluation of anthelmintic properties,assessed using faecal nematode egg counts,of New Zealand native flax (Phormium tenax)

AIM: To investigate the anthelmintic properties of New Zealand native flax (Phormium tenax) for cattle, using a faecal egg count reduction test (FECRT).

METHODS: Twenty-six heifer calves with high (>300 epg) faecal nematode egg counts (FEC) were paired into two groups and fed either chopped flax leaves ad libitum, or barley straw at an equivalent dry matter (DM) rate, from Day 0–7; all were supplemented with 1.5 kg/head/day cereal feed supplement. On Days 8–10, all heifers were fed a common diet of 3.5 kg DM/ head/day barley straw and 1.5 kg/head/day cereal feed supplement. Heifers were weighed and faecal samples were collected on Days —6, 0, 7, and 10; samples were analysed in triplicate for FEC. The nutritive value of the flax and straw was analysed.

RESULTS: Both the flax and straw had low dry matter digestibility (DMD) and protein content. Although the flax-fed claves grew more than the controls, this may have been related to gut fill. Total daily faecal egg output was similar for flax and straw groups on Day 7 (14.7 vs 15.0 x 10⁶ eggs/day, respectively) and Day 10 (14.9 vs 15.1 x 10⁶ eggs/day, respectively). There was no difference in the change in FEC with time between the calves fed flax or straw diets.

CONCLUSION: Consumption of flax leaves did not reduce FEC in calves with a mixed nematode infection.     

Effect of copper oxide wire particles dosage and feed supplement level on Haemonchus contortus infection in lambs

The objective of the experiment was to determine the optimal dose of copper oxide wire particles (COWPs) to reduce infection of Haemonchus contortus in male lambs. Five to six-month-old hair breed lambs were housed on concrete and fed 450 (L; n = 25) or 675 g (H; n = 25) corn/soybean meal supplement and bermudagrass hay. In July, lambs were inoculated with 10,000 L(3) larvae (97% H. contortus; Day 0). Lambs were administered 0, 2, 4, or 6 g COWP on Day 28. Concentrations of copper in the liver were determined. There were no effects of supplement level on concentrations of copper in the liver and a linear relationship existed between COWP treatment and concentrations of copper in liver (P < 0.001). Least squares means of the 0, 2, 4, 6 g COWP treatments were 62.2, 135.7, 161.1, and 208.4 ppm (P < 0.001). Between Days 0 and 28, PCV declined and by Day 42, PCV of all COWP-treated lambs was markedly higher than control lambs and remained higher (COWP x day, P < 0.05). By Day 21, PCV was greater in the H compared with the L group of lambs (P < 0.001). Within 14 days of COWP treatment FEC declined from more than 8000 eggs/g (epg) to less than 250 epg in all COWP-treated lambs (COWP x day, P < 0.001). The numbers of H. contortus in the abomasum were greatly reduced in all COWP-treated groups of lambs and remaining nematodes were predominantly males. FEC were greater in L versus H supplemented lambs and values decreased to a greater extent in H lambs when treated with COWP (supplement x COWP; P < 0.02). The 2 g COWP was effective in alleviating H. contortus infection and reducing number of egg-laying nematodes in the abomasum with the lowest concentration of copper in the liver of the COWP treatment groups. PCV values were more favorable for lambs fed the higher level of supplement, especially when FEC were greater than 8000 epg.     

Epidemiology of nematode parasites of sheep around Jimma,southwestern Ethiopia

An investigation was made into the epidemiology of nematode infections of sheep in two districts of Jimma zone, southwestern Ethiopia. We used two approaches—long-term monitoring of identified sheep for nematode infection and abattoir or market survey for analysis. In the first monitoring regime, we used 80 lambs [40 sheep (20 per sex) from each district (Dedo and Yebu)] averaging 4–5 months of age. Faecal egg counts (FEC), packed cell volume (PCV) and body weight changes were monitored over a period of 1 year. Additionally, faecal samples were collected (on a weekly basis) from sheep brought to abattoir/market for 1 year to monitor faecal egg counts. The nematode parasite burden, as judged by FEC and PCV, was generally low indicating that the climatic conditions are not conducive to the development and survival of nematode eggs and the free-living stages; hence, little transmission occurred. In the experimental flocks, the highest FEC and lower PCV were recorded during the long rainy season (June to September) with peak in August and September. Faecal samples collected from abattoir/market also followed the same trend. Results from experimental sheep indicated that location had a significant (P < 0.01) effect on FEC, PCV and average daily body weight gain. The FEC and PCV for sheep in Yebu (mid-altitude) district were 126 ± 3.33 and 30.6 ± 0.26, whereas the values for Dedo (highland) were 93 ± 4.35 and 32.0 ± 0.21, respectively. The results indicate that the highland areas are comparatively less favourable to the survival and development of nematodes. Female lambs had lower FEC and higher PCV compared to male lambs (P < 0.05). The overall nematode parasite challenge in the area, however, is low. We, therefore, recommend rotational grazing management combined with monitoring parasite load and selective treatment to reduce productivity loses and pasture contamination.     

Real-time quantitative polymerase chain reaction detection of haemoplasmas in healthy and unhealthy dogs from Central Macedonia, Greece

Objectives : The aims of this study were to determine the prevalence of canine haemoplasmas, Mycoplasma haemocanis and “Candidatus Mycoplasma haematoparvum” infection in Central Macedonia, Greece, and to evaluate any associations between canine haemoplasma infection and clinical presentation, selected laboratory data or the presence of ticks. Methods : Genomic DNA was purified from excess blood (n=151) submitted for haematological examination. Purified DNA was subjected to species‐specific quantitative polymerase chain reaction assays duplexed with a canine DNA control quantitative polymerase chain reaction. Clinical records were retrospectively examined and selected clinical parameters were compared to haemoplasma infection status. Results : Nine samples were excluded due to inadequate canine DNA polymerase chain reaction results. Of the remaining 142 samples: eight (5·6%) were positive for M. haemocanis alone, six (4·2%) were positive for “Ca. M. haematoparvum” alone and one (0·7%) was dual positive. No association was found between haemoplasma status and age, sex, breed, health status, presence of anaemia, selected biochemistry parameters, presence of ectoparasites, routine ectoparasiticide treatment or the presence of selected tick‐borne diseases.     

Relative resistance of Dorper crossbred ewes to gastrointestinal nematode infection compared with St. Croix and Katahdin ewes in the southeastern United States

A study was conducted to evaluate relative resistance of Dorper crossbred (DO), Katahdin (KA), St. Croix (SC), and Hampshire (HA) ewes to natural and experimental gastro-intestinal (GI) nematode infection over a 20-month period. The objective of Experiment 1 was to evaluate breeds for resistance to infection acquired naturally from mixed grass pastures. In Year 1 (May-December 2000) de-worming of ewes occurred during wet, hot conditions in July and during late pregnancy in December. In Year 2 (January-December 2001), ewes were de-wormed after fecal egg count (FEC) for a breed group rose above 1000 eggs per gram (epg) or after blood packed cell volume (PCV) of an individual ewe fell below 20. FEC was determined every 28 days and PCV every 14-28 days. In both the years, ewes were pastured together, except during the 28-days breeding periods, on tall fescue, bermudagrass, or ryegrass, and rotated among pastures dependent on forage availability. Ewes were in good or excellent condition (body condition score of 3-4 out of 5) throughout the study. The objective of Experiment 2 was to evaluate the breeds for relative resistance to an experimental infection with Haemonchus contortus infective larvae. Both PCV and FEC were determined every 7 days from 14 to 42 days after inoculation with 30000 infective larvae per ewe. In Experiment 1, Year 1, FEC was slightly greater and PCV was lower from July to September in DO ewes (breed x time, P<0.001). In Year 2, de-worming occurred 14 days later in DO ewes compared with other breed types. Otherwise PCV and FEC were similar among the hair breeds and higher and lower, respectively, compared with HA ewes (breed x time, P<0.001). In Experiment 2, FEC and PCV were similar among hair breeds; FEC was lower and PCV higher in hair breeds compared with that of HA ewes (P<0.01). Relative resistance of mature Dorper crossbred ewes was comparable to that of Katahdin and St. Croix ewes and superior to that of Hampshire ewes.     

Experimental Inoculation of Pregnant Swine with Type 1 Bovine Viral Diarrhoea Virus

The ability of bovine viral diarrhoea virus type 1 (BVDV‐1) to induce transplacental infection in pigs was evaluated. Control pigs (n = 4) were sham‐inoculated while infected pigs (n = 4) were intranasally inoculated with BVDV‐1 on day 65 of gestation. Blood samples were tested throughout the study for BVDV and antibody to BVDV. On day 110 of gestation, a Caesarean section was performed. Serum was obtained for virus isolation and antibody determination from all piglets, and all experimental animals were killed. Tissues were collected for virus isolation and histopathology. Bovine viral diarrhoea virus was isolated on days 5 and 7 after infection and seroconversion was demonstrated in all infected gilts; however, BVDV was only isolated from one fetus from an infected pig. Viraemia and seroconversion were demonstrated in the pregnant gilts; however, transplacental infection at day 65 of gestation in the pig was not consistently demonstrated.     

Changes in the body condition scores of Nigerian West African Dwarf sheep experimentally infected with mixed infections of Haemonchus contortus and Trichostrongylus colubriformis

Weekly changes in body condition scores and body weights were determined in West African Dwarf sheep following trickle exposure to weekly escalating mixed infections with Haemonchus contortus and Trichostrongylus colubriformis starting with single doses of 500 infective larvae (L(3)) at wk1, 1000 L(3) at wk2, 2000 L(3) at wk3 and 4000 L(3) at wk4. Thirty-three animals were randomly assigned to two groups namely, Group 1 (n=27) and Group 2 (n=6). Group 1 animals were used for experimental infections and Group 2 animals served as uninfected controls. Body condition scores (BCS), body weights (BW), faecal egg counts (FEC) per gram of faeces and worm burden (Wb) were determined using standard procedures. Lambs in group 1 with epg ≤ 1000 were classified as strong responders (n=16), those with epg between 1000 and 10,000 as intermediate (n=5) and lambs with epg>10,000 on any sampling day were classified as weak responders (n=6). Based on this classification, the mean epg of lambs classified as strong responders was significantly lower (P<0.001) than the epg of lambs classified as weak responders. The body condition scores and body weights of the strong responder lambs were comparable with those of the uninfected control and they were significantly higher than those of the weak and intermediate responders from wk5 till the end of the experiment. There was an inverse and significant relationship between the average BCS of days 56 and 59 and the average FEC of days 56 and 59 (r(s)=-572; P=0.017; n=27) as well as with Wb (r(s)=-531; P=0.028; n=27). We concluded that changes in the body scores are good indicators of the intensity of gastrointestinal nematode infection in the Nigerian WAD sheep.     

Effects of non-genetic factors on responses to gastro-intestinal nematode infections in Ethiopian sheep

The effects of non-genetic factors on responses to gastrointestinal nematode infections were examined in artificially infected indigenous Horro and Menz lambs in the central highlands of Ethiopia. The experiment involved lambs of two sexes, infection and monitoring regimens (3 levels), dam parity (4 levels) and birth types (2 levels). A total of 152 weaner lambs (103 Menz and 49 Horro) were used. Endoparasite challenge involved three infection periods (primary, secondary and tertiary) each separated by an anthelmintic treatment. Data were collected on faecal egg counts (FEC), packed cell volume (PCV), worm count, and weight gain. Female lambs were found to harbor lower numbers of T. colubriformis counts and had lower (p < 0.01) FEC in tertiary infection compared to males. Younger animals were more susceptible to the nematode infections than older animals as measured by all the parameters studied (p < 0.01). Twins had higher FEC in both secondary and tertiary infections compared to singles. Lambs born to dams of first parity had higher PCV and lower FEC values during the tertiary infection compared to other parities. These results would be used as basic information in designing a more rational control strategy and should also help to advise on better management care for less resistant sheep.     

Bovine viral diarrhoea virus (BVDV) subgenotypes in diagnostic laboratory accessions: distribution of BVDV1a, 1b, and 2a subgenotypes

The prevalence of bovine viral diarrhoea virus (BVDV) biotypes and subgenotypes was determined from 131 BVDV positive samples from a diagnostic laboratory. The majority of the isolates were from Oklahoma; however, other states including Kansas, Texas, and Arkansas were represented. These BVDV samples were from submissions of 76 live animals and 55 necropsy samples. There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with mixed NCP and CP biotypes. The NCP isolates were more common (P < 0.05) than the CP and NCP/CP combination. The BVDV samples were segregated into three subgenotypes by differential PCR and sequencing of a viral genomic region, 5'-untranslated region (5'-UTR). There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a, 34/131 (26.0%) (P < 0.05). The organ system involvement included the major categories such as respiratory, digestive, mixed/multiple organs, abortions, and persistent infections (PI). All three BVDV subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains present in U.S. vaccines.     

Validation of a bulk tank milk antibody ELISA to detect dairy herds likely infected with bovine viral diarrhoea virus in New Zealand

AIMS: To assess the sensitivity and specificity of a bulk tank milk (BTM) antibody enzyme-linked immunosorbent assay (ELISA) to detect likely infection of a dairy herd with bovine vi- ral diarrhoea virus (BVDV). The ELISA was subsequently used to estimate the prevalence of likely infected herds in parts of the North Island of New Zealand.

METHODS: BTM samples from 724 randomly selected dairy herds in the Waikato, Bay of Plenty and Northland regions of New Zealand were tested for BVDV antibodies. From this group, 20 herds were again randomly selected from each of the quartiles of the ELISA percentage inhibition (%INH) result. From each participant herd, serum from 15 randomly selected calves aged 6–18 months and 15 cows was collected and tested using an indirect blocking ELISA for BVDV antibodies.

RESULTS: Among serum results from calves from 50 herds available for analysis, 34 (68%) herds were classified as likely non-infected (0-3 seropositive among 15 calves) and 16 (32%) as likely infected (5–15 seropositive among 15 calves). Receiver- operator characteristic (ROC) analysis identified an optimal cut-off for BTM of 80%INH associated with 81% sensitivity and 91% specificity for likely herd infection. The prevalence of BVDV antibodies in cows within herds and %INH for BVDV in bulk milk were positively correlated (p<0.01). The association between bulk milk %INH and the prevalence of BVDV antibodies in calves was stronger than the same association in cows. Based on the threshold of 80%INH, the 95% confidence interval (CI) for prevalence of likely infection in the 724 herds in the Waikato, Bay of Plenty and Northland regions of New Zealand was 12–17%. Vaccination against BVDV was not significantly associated with the likely infection status of the herd based on prevalence of BVDV antibodies among calves.

CONCLUSION: An ELISA test result for BVDV antibodies in BTM ≥80%INH can be used as a threshold to indicate the presence of likely infection with BVDV in dairy herds in New Zealand, with 81% sensitivity and 91% specificity.     

Accuracy of the FAMACHA system for on-farm use by sheep and goat producers in the southeastern United States

FAMACHA is a practical on-farm system designed to provide small ruminant producers a tool for improving their management of Haemonchus contortus infections. Although this system has become very popular and widely accepted by small ruminant producers in many regions of the southern United States, there is very limited data reported on the effectiveness of the FAMACHA system when performed by farmers. The objective of this study was to evaluate the accuracy of the FAMACHA system for on-farm use by small ruminant producers during the summer season. Small ruminant producers from Georgia, Louisiana, Florida, and Puerto Rico were trained to use the FAMACHA system by veterinarians and scientists experienced with this method. FAMACHA scores were assigned at least every 2 weeks by producers to weaned and mature sheep (n=552) and goats (n=676) of various breeds and ages between April and September 2004. At intervals that varied among farms from 2 to 8 weeks, researchers determined body condition scores (BCS; 1=thin and 5=fat) and collected blood and feces from a group of animals selected randomly to determine packed cell volume (PCV) and fecal egg counts (FEC). Two separate anemia thresholds were evaluated; these were defined by either FAMACHA score (>or=3 versus >or=4) or PCV (or=3 were considered anemic and PCV cutoff was or=3 were considered anemic and PCV cutoff was 相似文献   

Helminths in horses: use of selective treatment for the control of strongyles

The current level of anthelmintic resistance in the horse-breeding industry is extremely high and therefore more emphasis is being placed on studies that focus on the judicious use of anthelmintic products. The aims of the study were to: 1) establish if there is variation in the egg excretion pattern of strongyles between the different age classes of Thoroughbred horses in the Western Cape Province (WCP), 2) test if a selective treatment approach successfully reduces the number of anthelmintic treatments and maintains acceptably low helminth burdens in adult Thoroughbred horses, and 3) evaluate the efficacy of subsampling large horse herds for faecal egg counts (FECs) to monitor the strongyle burden. In 2001 the FECs of 4 adult mare, 5 yearling and 3 weanling herds from 8 different farms were compared in the WCP Within the mare herds there were generally fewer egg-excreting individuals with lower mean FECs compared with the younger age classes. Individual faecal samples were collected every 3-4 weeks from 52 adult Thoroughbred mares from 1 farm in the WCP during a 12-month period (2002/2003). Animals with strongyle FECs > or =100 eggs per gram (epg ) were treated with an ivermectin-praziquantel combination drug (Equimax oral paste, Virbac). The mean monthly strongyle FEC for the entire group was <300 epg throughout the study and the number of treatments was reduced by 50 %. Resampling methods showed that an asymptote to mean FEC was reached at 55 animals for each of the pooled weanling, yearling and mare egg counts. Resampling within 4 different mare herds recorded asymptotes of between 24 and 28 animals. Subsampling entire herds for FECs therefore provided an effective approach to treatment management. This study demonstrates that selective treatment is both a practical and an effective approach to the management of anthelmintic resistance.     

Gastrointestinal nematode infections in small ruminants under the traditional husbandry system during the dry season in southern Ethiopia

A cross-sectional study was conducted from November 2008 to February 2009 to investigate the prevalence and intensity of infection and risk factors of gastrointestinal (GI) nematodes in small ruminants kept under the traditional husbandry system in two districts in southern Ethiopia. Faecal samples collected from a total of 510 small ruminants (284 sheep and 226 goats) and analysed by a modified McMaster technique revealed that 222 animals (43.5%) were found to be infected with one or more GI nematodes. Five identical genera of nematodes were found in both sheep and goats, which in order of predominance were Haemonchus (56.3%), Trichostrongylus (39.6%), Oesophagostomum (22.9%), Trichuris (21.6%) and Bunostomum (10.4%). No significant (p > 0.05) differences were observed between sheep and goats proportions except for Trichuris (p < 0.05). In both sheep and goats, most of the animals were heavily infected showing faecal egg counts (FECs) above 1,200 epg. Sheep had a significantly (p < 0.05) higher mean FEC than goats. In sheep and goats, both the prevalence of GI nematodes and mean FEC were significantly (p < 0.001) associated with body condition score and faecal consistency but not with district, sex and age (p > 0.05 for each factor). In conclusion, the observation of a strong association of GI nematodes with poor body condition coupled with heavy intensity of infection in the majority of infected animals and an abundance of nematode genera of widespread economic and pathological significance warrants the institution of appropriate control measures that should necessarily include improvement of the nutritional status of the animals.     

Selective and on-demand drenching of lambs: Impact on parasite populations and performance of lambs

AIM: To determine whether drenching regimes for lambs by which a proportion (10%) of the heaviest animals was selectively left untreated, or animals are only drenched ‘on demand’ when faecal nematode egg counts (FEC) exceeded a threshold level, would result in measurable increases in parasite larval challenge in the autumn and/or decreases in the performance of lambs.

METHODS: A replicated study compared three drenching strategies in which mobs of lambs (n=360 in total) received either: a five-drench preventive programme, administered to all animals (Treatment 1); a five-drench preventive programme, but the 10% heaviest animals left untreated each time (Treatment 2); or drench treatments administered only when FEC exceeded 500 eggs per gram of faeces (epg) (Treatment 3). After the five-drench programme, animals in Treatments 1 and 2 were treated according to FEC as for Treatment 3. A triple-combination drench containing ivermectin, oxfendazole and levamisole, administered orally, was used for all treatments. There were nine farmlets, allowing three replicates of each treatment, in a completely randomised design. Parasite infestations on pasture were measured in autumn by pasture plucks, and worm burdens were monitored in tracer lambs, while the performance of lambs was assessed by liveweight gains, fleece weights, and body condition and dag scores.

RESULTS: Increased numbers of Haemonchus contortus and Trichostrongylus colubriformis larvae on pasture were found in the autumn on farmlets treating selectively or on-demand (Treatments 2 and 3). No differences were detected in other parasite species. Mean liveweight gains did not differ between treatments but some differences were detected between drenched and undrenched lambs in Treatment 2. Mean body condition and mean dag scores of lambs in Treatment 3 tended to be lower and higher, respectively, than those of lambs in Treatment 1; Treatment 2 was generally intermediate.

CONCLUSIONS: Drenching strategies for lambs designed to slow the development of anthelmintic resistance, by increasing the pool of susceptible worms available to dilute resistant survivors after treatment, resulted in increased numbers of H. contortus and T. colubriformis but not other species of parasite on pasture. The increased parasite challenge to lambs in the autumn was associated with small production losses, which may be acceptable to farmers wishing to implement such strategies. It is clear that further work is required on the interaction between management practices and the population dynamics of parasites, especially with regard to creating pools of susceptible genotypes to slow the development of drench resistance.     

The effectiveness of copper oxide wire particles as an anthelmintic in pregnant ewes and safety to offspring

The objective of the experiment was to determine the effectiveness of copper oxide wire particles (COWP) in pregnant ewes and safety to lambs. COWP have been used recently as an anthelmintic in small ruminants to overcome problems associated with nematode resistance to chemical dewormers. Doses of COWP (相似文献   

Genotypic characterization of Chilean llama (Lama glama) and alpaca (Vicugna pacos) pestivirus isolates

Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities worldwide where they have been introduced. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus, in particular Bovine Viral Diarrhea (BVDV), but there is little data available on Pestivirus infections in SACs. In this study we aimed to detect and identify Pestivirus genotypes and subgroups infecting SACs in both wild and confined environments. Samples were collected from 136 llamas and 30 alpacas from different areas in the Chilean Altiplano (wild animals), and from 22 llamas and 26 alpacas diagnosed as Pestivirus positive from the Metropolitana region in Chile (confined animals). Seroneutralization tests showed titers lower than 2 in all 166 samples from Chilean Altiplano. These samples were also negative to BVDV isolation, indicating that these animals have not been exposed to Pestivirus. After reactivation of positive samples from the Metropolitana region, the 5′ non-codifying region (5′NCR) and E2 glycoprotein were amplified by RT-PCR from the Pestivirus genome. Viral sequences were pairwise compared and phylogenetic trees were constructed. The 5′NCR analysis showed that all 12 sequenced isolates belonged to BVDV-1. Of particular interest, isolates from eight llama and two alpaca were BVDV-1j and two alpacas were BVDV-1b. In agreement with these results, E2 phylogenetic analysis rendered a similar grouping indicating that all 16 isolates belong to BVDV-1. However, the lower availability of E2 sequences determines the creation of a smaller number of sub-groups than the 5′NCR sequences. Based on the E2 sequences, the 5′NCR BVDV 1j group consisting of all the llamas and 3 alpacas are completely included in the E2 BVDV 1e group. Due to the universal availability of the 5′NCR segment, we propose the classification of these Chilean llamas and alpacas Pestivirus isolates as BVDV 1j and BVDV 1b respectively. Thus, this is the first time BVDV-1j is obtained in SACs. In addition, these results indicate Pestivirus infection in llamas and alpacas is associated with bovine population as genotypes and sub-groups are the same as those affecting Chilean livestock.     

Epidemiology and genetic characterization of BVDV,BHV-1, BHV-4, BHV-5 and Brucella spp. infections in cattle in Turkey

The aim of the study was to determine the epidemiological data of bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine herpesvirus-4 (BHV-4), bovine herpesvirus-5 (BHV-5) and Brucella–associated cattle that were previously reported to have abortion and infertility problems in Ankara, Corum, Kirikkale and Yozgat provinces, Turkey. Whole blood and sera samples were obtained from 656 cattle, and antibodies against Brucella spp. were detected in 45 (6.86%) and 41 (6.25%) animals by Rose Bengal plate and serum tube agglutination tests, respectively. The seropositivity rates against BVDV, BHV-1 and BHV-4 were 70.89%, 41.3% and 28.78%, respectively. RT-PCR and PCR were performed to detect RNA and DNA viruses in blood samples, respectively. The BVDV 5′-untranslated region and BHV-1 gB gene detected in this study were phylogenetically analyzed. The BVDV strains analyzed in this study were closely related to those previously reported from Turkey. The nucleotide sequence from the BHV-1 strain detected in this study is the first nucleotide sequence of BHV-1 circulating in this area of Turkey deposited in the GenBank. The presence of Brucella spp. and prevalence of BHV-1, BHV-4 and BVDV in cattle should be further investigated throughout these regions.     

Multiple diagnostic tests to identify cattle with Bovine viral diarrhea virus and duration of positive test results in persistently infected cattle

Several tests for Bovine viral diarrhea virus (BVDV) were applied to samples collected monthly from December 20, 2005, through November 27, 2006 (day 0 to day 342) from 12 persistently infected (PI) cattle with BVDV subtypes found in US cattle: BVDV-1a, BVDV-1b, and BVDV-2a. The samples included clotted blood for serum, nasal swabs, and fresh and formalin-fixed ear notches. The tests were as follows: titration of infectious virus in serum and nasal swabs; antigen-capture (AC) enzyme-linked immunosorbent assay (ELISA), or ACE, on serum, nasal swabs, and fresh ear notches; gel-based polymerase chain reaction (PCR) testing of serum, nasal swabs, and fresh ear notches; immunohistochemical (IHC) testing of formalin-fixed ear notches; and serologic testing for BVDV antibodies in serum. Of the 12 animals starting the study, 3 died with mucosal disease. The ACE and IHC tests on ear notches had positive results throughout the study, as did the ACE and PCR tests on serum. There was detectable virus in nasal swabs from all the cattle throughout the study except for a few samples that were toxic to cell cultures. The serum had a virus titer ≥ log₁₀ 1.60 in all samples from all the cattle except for 3 collections from 1 animal. Although there were several equivocal results, the PCR test most often had positive results. The BVDV antibodies were due to vaccination or exposure to heterologous strains and did not appear to interfere with any BVDV test. These findings illustrate that PI cattle may be identified by several tests, but differentiation of PI cattle from cattle with acute BVDV infection requires additional testing, especially of blood samples and nasal swabs positive on initial testing. Also, calves PI with BVDV are continual shedders of infectious virus, as shown by the infectivity of nasal swabs over the 11-mo study.