The Effect of Management Factors on the Seroprevalence of Anaplasma marginale in Bos indicus cattle in the Mexican Tropics

A cross-sectional study with a two-stage design and proportional distribution was carried out to determine the effect of management factors on the seroprevalence of Anaplasma marginale in Bos indicus cattle in the Mexican tropics. Serum was obtained from 384 cattle aged 1-2 years on 92 farms. The number of samples was proportionally distributed according to the number of farms in eastern Yucatan. Antibody activity against A. marginale was assessed using the card agglutination test. A primary screening using a 2 x K contingency table of the exposed variables with the outcomes was performed. All variables for which p < 0.20 were included in a fixed-effects log regression. The seroprevalence in the cattle was 69.75% (SE +/- 0.02). Sixty-four per cent of the farms had a seroprevalence > or = 75%. The risks related to managemental factors were stocking density ( > or = 1 animal/ha, OR = 10.94), type of acaricide (pyrethroids, OR = 3.8), dipping interval ( > 60 days, OR = 0.13) and type of veterinary instruments used (needles, scalpels, ear tattoos, and dehorners, OR = 0.17).     

In vivo anthelmintic activity of Phytolacca icosandra against Haemonchus contortus in goats

Two separate controlled and blinded studies were conducted to confirm the dose and non-interference of spinosad and milbemycin oxime (MO) administered orally in combination or alone to dogs for the treatment and control of experimentally induced flea infestations (Ctenocephalides felis) and adult hookworm infections (Ancylostoma caninum). For each study, dogs were allocated randomly based on pre-treatment adult flea and hookworm egg counts to one of four treatment groups of 10 animals each. In each study, spinosad and MO in combination, using the lower half (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial dose band (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) of each active ingredient, or individually alone using the full dose range, were given orally to dogs on Day 0 using a tablet formulation. A placebo control was treated similarly. In one study, on Days -1, 5, 12, 19, 28 and 35 each dog was infested with approximately 100 unfed adult C. felis obtained from the investigator's established flea colony. All dogs were infested via the same method. Forty-eight hour post-infestation flea comb counts were conducted on Days 1, 7, 14, 21, 30 and 37 and were used to determine the knockdown and residual flea activity. In the second study, on Day -27 each of 48 dogs were experimentally inoculated with 100 third-stage infective larvae of the hookworm, A. caninum. Dogs were treated on Day 0 and necropsied on Day 7 or Day 8. All nematodes in the intestinal tract were collected on Day 7 or Day 8, identified and counted by species and stage. Post-treatment, the geometric mean live flea counts were significantly different (p-value<0.0001) between the spinosad/MO combination and the spinosad only treatment groups as compared to the vehicle control group. The flea counts in the MO only group and the control group were not statistically different. The spinosad and MO combination group and the spinosad only treatment group demonstrated significantly different knockdown (100%) and post-treatment residual flea efficacy at Day 30 was 100% for both groups as compared to the vehicle control. The presence of MO in combination with spinosad did not interfere with the flea efficacy of spinosad as compared to the spinosad only group. MO alone did not demonstrate any flea efficacy. Post-treatment, the geometric mean A. caninum worm counts were significantly different (p-value<0.0001) between the spinosad and MO combination group as compared to the vehicle control group. The worm counts in the MO only group and the combination group were not statistically different. The spinosad and MO combination group (99.8% reduction) and the MO only treatment group (99.5% reduction) both demonstrated significantly different hookworm efficacy as compared to the vehicle control group. The presence of spinosad in combination with MO did not interfere with the hookworm efficacy of MO as compared to the MO only group. Spinosad alone did not demonstrate any hookworm efficacy. In summary, flavored spinosad and MO combination tablets administered orally to dogs at the lower end (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial tablet unit dose range (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) were both safe and highly efficacious delivering 100% knockdown and 30 days of residual adult flea control on experimentally infested dogs as well as >99% adult hookworm efficacy evaluated under laboratory conditions. Interference between either drugs was not demonstrated for both of these dose limiting parasites.     

Prevalence of benzimidazole resistant nematodes in sheep flocks in Yucatan,Mexico

Thirty-eight sheep flocks, located in three municipalities in the Eastern Yucatan, Mexico, were surveyed for gastrointestinal nematodes resistant to benzimidazole (BZD) anthelmintics (AH). On each flock, 30 sheep were randomly distributed into two groups of 15 animals: albendazole group (5mg/kg BW) and untreated control group. Animals were refrained from any food (either browsing/grazing or supplement) for a period of 16 h prior to treatment. Faecal egg counts (FEC) and larval cultures were performed 10 days after anthelmintic treatment. Percentage reduction and 95% confidence intervals were determined. Anthelmintic resistance (AR) was declared when the percentage reduction in FEC was <95% and the 95% confidence interval was <90%. AR was suspected when only one of the two criteria was met. The survey indicated that AR occurred in 15.8% (n=6) (95% confidence interval=+/-11.6%) and was suspected in 23.7% of the farms (n=9) (95% confidence interval=+/-13.3%). Post-treatment larval cultures indicated that Haemonchus was the only resistant genus. The questionnaire survey showed that most farmers (92%) considered their sheep a secondary activity to cattle production. The majority of farmers (97.4%) treat their animals according to visual appreciation of weight. However, most farmers (79%) treat their flocks at very low frequencies (<3 times per year). Drug rotation was performed every 12 months or more by 84.2% of farmers. Anthelmintics used were: macrociclic lactones (47.4%), BZD (39.5%), levamisol (10.5%) and 1 farmer used closantel.     

In vitro acaricidal effect of tannin-rich plants against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

The objectives of this study were to evaluate the in vitro acaricidal effects of lyophilized extracts of four tannin rich plants (Acacia pennatula, Piscidia piscipula, Leucaena leucocephala and Lysiloma latisiliquum) against diverse stages of Rhipicephalus (Boophilus) microplus, and to asses whether tannins were involved in the acaricidal effect using polyethylene glycol (PEG) to block tannins. Larval immersion (LIT) and adult immersion (AIT) tests were used to evaluate the acaricidal effect of each of the lyophilized extracts against larval and adult stages of R. microplus respectively. Larvae and adult ticks were exposed to increasing concentrations of each plant extract (0, 1200, 2400, 4800, 9600 and 19,200 μg ml(-1)) for 10 min. Larval mortality was recorded at 48 h post-incubation. Adult mortality was recorded daily over 14 days, at which point their reproductive efficiency was evaluated. PEG was added to the extracts to verify whether tannins were involved in the acaricidal effect. The effect on egg laying inhibition and larval mortality was analyzed using the GLM procedure in SAS. A Kruskal-Wallis test was used to assess the effect of PEG on LIT results. Calculation of the lethal concentration 50 (LC50) was performed using a probit analysis. All extracts reduced the viability of R. microplus larval stages (P<0.001), and viability was restored with the addition of PEG suggesting an important role of tannins in the acaricidal effect (P<0.001). The LC50 values of L. latisiliquum and P. piscipula plant extracts were 6.402 and 2.466 μg ml(-1). None of the tannin-rich plant extracts affected adult mortality (P>0.05). Lysiloma latisiliquum extract inhibited egg hatching of R. microplus (P<0.01). Tannin-rich plant extracts from A. pennatula, P. piscipula, L. leucocephala and L. latisiliquum showed potential acaricidal activity. Further in vivo studies are needed to confirm this finding.     

Prevalence and potential risk factors for amitraz resistance in Boophilus microplus ticks in cattle farms in the State of Yucatan, Mexico

Southern cattle tick, Boophilus microplus, control in the Mexican tropics is based on the application by spray, dip or pour-on of acaricides. In Mexico the first report of ticks resistant to amitraz was reported in 2002. Amitraz resistance affecting the cattle industry of Yucatan State, Mexico, has not been investigated. A cross-sectional study was carried out in order to investigate the possible association between management risk factors and the prevalence of B. microplus ticks resistant to amitraz in Yucatan, Mexico. Ninety-eight cattle farms were randomly selected and a sample of 20–30 engorged B. microplus females was collected from at least 10 cattle from each farm. Collected ticks were taken to the laboratory where the larval progeny were obtained. The modified larval immersion test was used to detect ticks resistant to amitraz. A discriminating dose of 0.0002% amitraz was used. A questionnaire was applied on each collaborating farm. For the statistical analysis a 2 × 2 contingency table was used to identify potential risk factors. The prevalence of farms with B. microplus resistant to amitraz was 19.40%. However, even though tick resistance to amitraz is a present problem there was no evidence that it was associated with the risk factors analyzed in this survey.     

Efficacy of moxidectin 0.5% pour-on against naturally acquired nematode infections in cattle in the Mexican tropics

The efficacy of 0.5% moxidectin pour-on in cows with naturally acquired nematode infections was evaluated. The study was carried out in a ranch in Veracruz, Mexico. Four groups of 15 cows were randomly allocated. Animals in the treated group received 0.5% moxidectin pour-on at a dose of 0.5 mg/kg body weight on a single occasion. The other two groups remained as untreated controls. Fecal samples from all cattle were taken on days 0 (pre-treatment), 7, 14, 28 and 60 (post-treatment, PT). Fecal egg-counts were determined using a modified McMaster technique and fecal cultures were performed to identify gastrointestinal nematodes infected larvae (L(3)). Treatment with moxidectin was associated with a significant reduction in fecal trichostrongyle egg-counts compared with the controls; efficacy was 100% at 28 days PT. Haemonchus spp. and Strongyloides spp. were the two genera identified from coprocultures.     

Management factors associated with Babesia bovis seroprevalence in cattle from eastern Yucatán, Mexico.

The effect of management on the seroprevalence of Babesia bovis was studied in 399 Bos indicus cattle 1–2 years old from 92 farms in the eastern Yucatán, México. The management factors studied were: farm-type, production system, herd size, farm size, stocking density, vector control, dipping interval, type of dipping, type of acaricide and cattle introduction to the farm. A cross-sectional study was carried out (2-stage cluster sampling). The number of serum samples was proportionally distributed according to the number of farms in the nine locations of eastern Yucatán, México (399 animals from 92 farms). Antibody activity to B. bovis was tested using an indirect ELISA. The farms with a seroprevalence ≤75% were considered as cases and those with seroprevalence >75% were considered as controls. The variables with p ≤ 0.20 were included in fixed effects logistic regression. The seroprevalence of the zone was 73.8% (66.3–81.3%). The following risk factors were found: Stocking density (<1 head/ha, OR = 4.04, CI (OR) = 1.20–13.62) and dipping interval (>60 days, OR = 5.07 CI (OR) = 1.26–20.48).     

Management factors associated with Babesia bovis seroprevalence in cattle from eastern Yucatán, Mexico

The effect of management on the seroprevalence of Babesia bovis was studied in 399 Bos indicus cattle 1–2 years old from 92 farms in the eastern Yucatán, México. The management factors studied were: farm-type, production system, herd size, farm size, stocking density, vector control, dipping interval, type of dipping, type of acaricide and cattle introduction to the farm. A cross-sectional study was carried out (2-stage cluster sampling). The number of serum samples was proportionally distributed according to the number of farms in the nine locations of eastern Yucatán, México (399 animals from 92 farms). Antibody activity to B. bovis was tested using an indirect ELISA. The farms with a seroprevalence ≤75% were considered as cases and those with seroprevalence >75% were considered as controls. The variables with p ≤ 0.20 were included in fixed effects logistic regression. The seroprevalence of the zone was 73.8% (66.3–81.3%). The following risk factors were found: Stocking density (<1 head/ha, OR = 4.04, CI (OR) = 1.20–13.62) and dipping interval (>60 days, OR = 5.07 CI (OR) = 1.26–20.48).     

Detection of different Leishmania spp. and Trypanosoma cruzi antibodies in cats from the Yucatan Peninsula (Mexico) using an iron superoxide dismutase excreted as antigen

Although human leishmaniasis has been reported in 20 states in Mexico, no case of leishmaniasis has been reported in cats to date. In the Yucatan Peninsula, it has been found that dogs may act as reservoirs for at least three Leishmania species (Leishmania mexicana, Leishmania braziliensis, and Leishmania panamensis). In this study we identified specific antibodies against these three Leishmania spp. and Trypanosoma cruzi in the sera from 95 cats from two States on the Yucatan Peninsula, namely Quintana Roo and Yucatan, by ELISA and Western blot techniques using whole extract and an iron superoxide dismutase excreted by the parasites as antigens. As well as demonstrating the presence of trypanosomatid antibodies in the feline population on the Yucatan Peninsula, we were also able to confirm the high sensitivity and specificity of the iron superoxide dismutase antigen secreted by them, which may prove to be very useful in epidemiological studies.     

Prevalence of cattle herds with ivermectin resistant nematodes in the hot sub-humid tropics of Mexico

The objective was to determine the prevalence of semi-intensive Bos indicus and Bos indicus × Bos taurus cattle herds with ivermectin (IVM) resistant nematodes in a sub-humid tropical zone of Mexico using the faecal egg count reduction test (FECRT). Thirty-three herds (28 beef and 5 dual purpose herds) were monitored in a period of 6 months (September 2008 to February, 2009). Only 14 of the 33 herds were included in the trial. The other herds had not enough animals with sufficient nematode eggs per gram of faeces (EPG) to be included in a FECRT. Some farms were visited twice trying to find more animals with egg counts higher than 150 EPG. In the 14 surveyed herds the calves were randomly distributed into two groups: (a) treatment group received 0.2mg of IVM/kg BW sc on day 0, and (b) control group without treatment. Faecal samples were obtained from each animal on days 0 and 14 post-treatment. Reduction percentages (% R) and 95% CI were calculated. The prevalence of cattle herds with IVM resistant nematodes was 78.6%. Those suspected of IVM resistance were 21.4%. All surveyed herds used IVM from two to three times a year (mainly beginning and end of the wet season) during 1-11 consecutive years. The farm with stronger resistance used IVM for 11 consecutive years (% R=0%; 95% CI=0-47%). Genera of nematodes resistant to IVM were: Ostertagia, Haemonchus, Cooperia and Trichostrongylus. A considerable effort is needed to perform FECRT in cattle herds under hot sub-humid tropical conditions.