Resistance to benzimidazole and macrocyclic lactone anthelmintics in cattle nematodes in Argentina.

In April 2003, persistent scouring and ill-thrift that was reported in calves form an intensive beef rearing operation in central Argentina despite treatments with benzimidazole and ivermectin. In order to conduct a controlled faecal egg count reduction test on this herd, 40 calves 5-8-months-old were selected on the basis that they had a nematode eggs per gram (epg) of faeces count greater than 150. Animals were divided into four groups (1-4) of 10 calves. Calves of groups 1-3 were treated, respectively, with subcutaneous injection of ivermectin (200 mcg/kg), ricobendazole (4 mg/kg) and levamisole (7.5 mg/kg), while calves of group 4 remained as untreated controls. The egg count reductions carried out 10 days later were lower than 15% in calves treated with ivermectin and ricobendazole, but 100% in animals receiving levamisole. Pooled post-treatment faecal cultures showed larval percentages of 92 and 95 for Haemonchus and 8 and 5 for Cooperia in the faeces of calves treated with ivermectin and ricobendazole, respectively. This is the first reported case of Haemonchus parasiting cattle showing simultaneous resistance to avermectins and benzimidazole type anthelmintics.

Abundance of stable flies on heifers treated for control of horn flies with organophosphate impregnated ear tags.

Ear tags containing 40% organophosphate insecticides (diazinon or diazinon plus chlorpyrifos-ethyl) were applied to control Haematobia irritans (L.) (Diptera: Muscidae) in treated (TG01 and TG02) and untreated (UG01 and UG02) groups of Holstein heifers born in 2001 and 2002, respectively. Control and treated groups were assessed for the abundance of Stomoxys calcitrans (L.) (Diptera: Muscidae) from August 2001 to April 2002 and again from August 2002 to April 2003. The treatment had a high efficacy for control of horn flies (maximum median number per heifer of TG01 and TG02 = 5) but a low effect on the abundance of stable flies. The total numbers of S. calcitrans were 1251 (42.9% of the total) and 1668 (57.1%) for TG01 and UG01, and 1423 (48.8%) and 1494 (51.2%) in TG02 and UG02, respectively. No significant difference in stable fly burden was found in 55 of the 76 weeks evaluated. A unimodal peak of abundance in the spring was found during the first fly season, and a bimodal abundance, with peaks in the spring and autumn, during the second season. No strong associations between horn fly and stable fly burdens was found in individuals of the CG01 (correlation coefficient = 0.13, P > 0.05) or CG02 (correlation coefficient = 0.538, P < 0.05, determination coefficient = 0.289).

A study of relative horn fly, Haematobia irritans (Diptera: Muscidae), abundance on Holstein steers and steers of two Holstein crosses.

A study was performed to determine if the number of horn fly (Haematobia irritans) adults differ significantly on Holstein (black and white coat color), Holstein x Holstein Friesian (black and white coat color) and Holstein x Jersey (black coat color) steers, 10-12 months old at the onset of the study. All steers were run together on lucerne paddocks and the number of flies counted at 30-day intervals from September 2000 to August 2001. No significant differences (P > 0.05, test of Kruskal-Wallis) were found in fly numbers, even in the period April-May 2001 when the infestation reached its peak. We were unable to demonstrate that coat color influenced horn fly abundance in the present study. It appears that none of the biotypes evaluated had any advantage for natural control of H. irritans.

Dynamics of cypermethrin resistance in the field in the horn fly, Haematobia irritans.

The toxicity of cypermethrin to the horn fly Haematobia irritans (L.) (Diptera: Muscidae) was determined for samples collected from untreated herds at a farm in central Argentina from October 1997 to May 2001. Field tests of the efficacy of cypermethrin against horn flies were first carried out at this farm in 1993, when the fly was shown to be susceptible to pyrethroids. Subsequently the horn fly populations on this farm were shown to have become resistant and, since 1997, the use of cypermethrin has been restricted to experimental purposes. In this study, fly samples collected in 1999, 2000 and 2001 were subjected to a polymerase chain reaction (PCR) to detect the presence of a specific nucleotide substitution in the sodium channel gene sequence, which has been associated with target site insensitivity to pyrethroids. This analysis showed that the level of cypermethrin resistance had diminished between 1997 and 2001. However, this was not sufficient to restore the efficacy of this pyrethroid to the level found prior to the onset of resistance. Heterozygous and homozygous resistant flies were detected in all samples of flies subjected to PCR diagnosis of alleles conferring target site resistance.

Toxicity of cypermethrin and diazinon to Haematobia irritans (Diptera: Muscidae) in its American southern range.

Synthetic pyrethroids are considered currently less effective than organophosphate insecticides for controlling Haematobia irritans on cattle in its American southern range. To test the validity of this hypothesis, horn fly populations from 95 cattle herds in central-northern Argentina and southern Brazil were evaluated to determine 50% lethal concentrations (LC(50)) to cypermethrin and diazinon, respectively. Horn flies were exposed to filter papers and glass vials treated with cypermethrin and diazinon, respectively, and LC(50) values were determined using probit analysis. Two and 78 fly populations showed LC(50) values indicating susceptibility and resistance to cypermethrin, respectively, in comparison to historical LC(50) values obtained when the horn fly populations were susceptible to cypermethrin. Five fly populations had an undetermined resistance status to cypermethrin. The diazinon tests showed no obvious indications of H. irritans resistance. Thirty nine percent of farmers still rely on pyrethroids to control horn fly populations on cattle which represents a decreasing proportion from a few years ago when horn fly control was carried out using pyrethroids almost exclusively. Organophosphates could provide control for horn flies resistant to pyrethroids; however, the reliance on chemicals alone to control H. irritans is not considered a proper approach for the future.

Long-term study of Haematobia irritans (Diptera: Muscidae) seasonal distribution in central Argentina with focus on winter fly abundance.

The seasonal distribution of Haematobia irritans (Linnaeus, 1758) was evaluated at 31 degrees 12' S-61 degrees 29' W, Santa Fe, Argentina from November 1992 to August 2000 by weekly fly counts on 20 Holstein cows not treated against ectoparasites. The analysis was mainly focused on winter fly abundance. Two peaks of abundance were found from spring to autumn. Adults of H. irritans were consistently found on cattle during winter, with increasing numbers from the end of July to late August. The only climatic parameter soundly correlated with the four week period, before each winter fly count, was the mean air temperature with the exception of year 1998. No significant relationship was found between level of cow infestation and relative humidity, saturation deficit and rainfall. Horn fly infestation on cows was lower than 20% from end of June to end of August only in 1995, when mean air temperature was consistently lower than 11 degrees C during the four week period previous to most fly counts. Conversely, the mean temperature was higher than 12 degrees C previous to fly counts in 1998, when most cows remained infested. The results indicate that a proportion (unknown) of immature stages of H. irritans were insensitive to diapause inducing factors and developed through winter.

Cattle breed-variation in infestation by the horn fly Haematobia irritans.

A study was carried out to assess the resistance of pure and cross-bred groups of cattle to the horn fly Haematobia irritans (Linnaeus) (Diptera: Muscidae) in northern Argentina. Pure-bred cattle were Criolla, Iberian Bos taurus Linnaeus (Artiodactyla: Bovidae) and Nellore, Bos indicus Linnaeus (Artiodactyla: Bovidae). Cross-bred cattle were Hereford, British B. taurus (34%) X Nellore (66%) and Hereford (66%) X Nellore (34%). All were heifers and animals were maintained in two groups, each containing a mixture of pure and cross-breeds. The lowest fly numbers were found on Criolla heifers and the highest on Hereford X Nellore cross-breeds. However, it could not be determined from this study whether this was a consequence of breed and/or size, as Criolla heifers were lighter than the corresponding Hereford X Nellore heifers. Fly numbers on the heifers followed an approximately negative binomial distribution. However, the ranking of individual animals in their level of infestation within subgroups was not consistent. Hence, culling the most infested heifers on any given date would at best give only a small improvement in H. irritans control.

Chlorfenapyr ear tags to control Haematobia irritans (L.) (Diptera: Muscidae) on cattle.

The efficacy of ear tags containing 30% chlorfenapyr (total tag weight=13g) to control natural Haematobia irritans (L.) infestations was evaluated for Holstein heifers in Rafaela, province of Santa Fe, Argentina. A group of heifers (TG) was treated with two ear tags (one tag per ear). A control group (ACG) was maintained in a paddock adjacent to the TG paddock and, a distant control group (DCG) was maintained 700m apart from the other groups. From day 4 to day 98 after treatment, H. irritans infestations of ACG were significantly higher (P<0.05, test of Kruskal-Wallis) than the corresponding infestation of TG, but significantly lower than infestation in DCG, probably due to the proximity of TG. The chlorfenapyr control period, with an efficacy higher than 90% to reduce horn fly populations, lasted for 9 weeks when TG infestation was compared to fly numbers in ACG, but increased to 12 weeks in comparison to DCG. The results of this study show that ear tags impregnated with chlorfenapyr are a useful alternative to pyrethroids and organo-phosphate compounds for horn fly control.

Evaluation of insecticide ear tags containing ethion for control of pyrethroid resistant Haematobia irritans (L.) on dairy cattle.

A field study was conducted in central Argentina to evaluate the efficacy of ear tags containing 36% ethion against pyrethroid resistant populations of Haematobia irritans on grazing dairy cattle. The treated group consisted of 45 milking Holstein cows which received two tags per head and the control consisted of 22 dry cows from the same cohort. Treated and control groups were grazed on similar lucerne pastures separated for a minimum distance of 800 m but they were not isolated from other cattle herds on the same or contiguous properties. In both groups, horn fly estimation were made weekly by examining cattle in the pastures with the aid of binoculars. The percentage reduction of fly numbers on treated cows was considered as efficacy of control provided by the tags. The ethion ear tags provided a range of 85-99% reduction in horn fly numbers for 16 weeks. During this period, the weekly mean number of H. irritans on the tagged cows ranged between 0.44 and 28.26 compared with 143.5 and 239.1 in control animals. The ethion ear tags could be a useful tool for the control of H. irritans mainly in areas where populations of this insect have developed resistance to pyrethroid insecticides.

Moxidectin pour-on for control of natural populations of the cattle tick Boophilus microplus (Acarina: Ixodidae).

Fifty Bos taurus x Bos indicus heifers naturally infested with Boophilus microplus ticks were divided into two groups of 25 heifers each. Individuals of one group were treated with moxidectin 0.5% pour-on at a dosage of 500 microg of moxidectin/kg body weight and heifers from the other group remained as untreated controls. An efficacy higher than 95% was found on days 7-21 after treatment by using female ticks 4.5-8.0 mm long as the main infestation parameter. A lower, but significant efficacy (p < 0.05) was also found on days 1 (32.3% efficacy) and 27 (73.4% efficacy) post-treatment. Significantly (p < 0.05) lower numbers of immature ticks were also observed on heifers of the treated group from days 7 through 27 after treatment. A lower engorgement weight of female ticks from treated heifers was found on days 1 and 21 after treatment. Treatment also affected reproductive performance (oviposition, egg hatch and number of eggs laid) of female ticks collected on Day 1.

Persistent activity of doramectin and ivermectin in the prevention of cutaneous myiasis in cattle experimentally infested with Cochliomyia hominivorax.

A study was conducted to evaluate the activity of a single administration of doramectin or ivermectin against severe, induced infestations of Cochliomyia hominivorax. Twenty-four Holstein bull calves were allocated to four groups of six animals each and treated either with saline, doramectin 1%, or either one of two formulations of ivermectin 1% at a dose rate of 200 microg/kg. On Day 12 after treatment, each calf was anesthetized and two wounds were created on the left side of the shoulder and rump of each calf and 2 h later, each wound was implanted with 100 newly hatched larvae of C. hominivorax. On Day 15 after treatment, the procedure was repeated on the right side of each calf. Wounds were examined daily for 5 days and evidence of live larvae was recorded. Doramectin provided reduction in myiasis of 90.9 and 83.3% at 12 and 15 days after treatment, respectively, compared to the saline control treatment (P < 0.0001). In contrast, there were no significant differences in the number of calves with myiasis between those treated with either of the ivermectin formulations and the saline control.

Skin lesions and cattle hide damage from Haematobia irritans infestations.

The horn fly Haematobia irritans L. (Diptera: Muscidae) has recently spread to Argentina and Uruguay and is believed to cause damage to cattle hides. Four groups of ten Holstein steers each were maintained for 58 weeks under different infestation levels with H. irritans to determine if it was the cause of this problem. Hides (chrome tanned) from steers maintained under minimum infestation level had 4.7 +/- 3.8% of the area damaged. Maintaining the steers under low H. irritans level for the last 44 days of the trial using insecticidal ear-tags, resulted in 29.5 +/- 15.8% of hide area being damaged. Steers that were treated with 5% cypermethrin pour-on, when the H. irritans population was close to 50 flies, showed that 31.3 +/- 16.6% of hide area was injured, and 46.6 +/- 12.8% of damaged hide area was found in hides from non-treated steers. Significant differences were found between mean hide damage from steers maintained continuously under low H. irritans infestation levels and all other groups. Hyperaemia was significantly lower in the skin of steers under low H. irritans infestation level than in the skins of non-treated steers and steers maintained under low-level infestations for the final 44 days. Eosinophil and mononuclear cell infiltration was significantly lower when the population of H. irritans was less than six per steer than when the population was more than 100 flies per steer. Low numbers of Stomoxys calcitrans were found in all groups, but most hide damage was presumed due to H. irritans.

Cypermethrin pour-on synergized with piperonyl butoxide: effects on Haematobia irritans (Diptera: Muscidae) natural populations resistant to cypermethrin.

Development of pyrethroid resistance in Haematobia irritans in Santa Fe province, Argentina, resulted in an increased use of pyrethroid insecticides, probably due to lack of suitable alternative treatments. We explored the efficacy of mixtures of cypermethrin and piperonyl butoxide (PBO) against pyrethroid-resistant H. irritans. Groups of 25 Holstein cows each, naturally infested with cypermethrin resistant H. irritans were assigned to treated or control groups in April, September, October and December 1997. Cattle in treated groups were medicated with pour-on oil formulations of 5% cypermethrin (dose = 4 mg per kg of body weight) with 5% or 10% PBO in April, and with a mixture containing 5% of both components thereafter. Efficacy was tested for 21 days after treatment. A treatment of 5% cypermethrin pour-on without PBO was evaluated in October 1997. Samples of horn flies were obtained before September, October and December treatments and exposed for 2 h to filter papers impregnated with different cypermethrin concentrations to determine the 50% lethal concentration (LC50). No difference in efficacy was found between cypermethrin pour-on formulations with 5% or 10% of PBO (more than 94% efficacy on day 21 after treatment). Efficacy of 5 % cypermethrin-5% PBO mixture decreased rapidly in the successive treatments (less than 40% efficacy was observed on day 21 after the December treatment), and the period after treatment with an efficacy higher than 95% was 14 days for the treatment carried out in April, 10 days in September; 7 days for the treatment performed in October and 4 days for the December treatment. The LC50 of cypermethrin was 36.6 microg per cm2 in September and increased to 116.6 and 226.1 microg per cm2 in October and December, respectively. It is concluded that the addition of PBO to cypermethrin did not provide a treatment that would give a long term control of pyrethroid resistant-horn flies.

Evaluation of injectable abamectin to control natural infestations of Haematobia irritans (Diptera: Muscidae) in cattle.

The effect of 200 micrograms/kg of body weight of injectable 1% abamectin to control Hematobia irritans (L.) was evaluated in heifers treated every 45 or 63 d versus control heifers. Fly numbers were estimated every 7 d with additional counts on days 1, 4, and 10 after treatment. Samples of feces were obtained up to 28 d after treatment from treated and control heifers to culture H. irritans in the laboratory. The H. irritans population reduction for 14 d after treatment ranged from 51% (3rd treatment of heifers treated every 45 d) to > 73% in the 1st treatments of both groups of heifers and the 2nd treatment of the group treated every 63 d. Medication every 45 d did not provide a better control than treatment every 63 d. The abamectin arrested the immature stages for 7 d after the 1st treatment; this period increased to 28 d after the 3rd treatment but its effect on heifer infestation was low, probably because of fly immigration. Abamectin may help maintain H. irritans populations below the threshold of economic damage.

Efficacy of dicyclanil in the prevention of screwworm infestation (Cochliomyia hominivorax) in cattle castration wounds.

A field trial was carried out during summer 1996 in a commercial beef cattle farm located in the central area of Argentina (30 degrees 16'S, 60 degrees 30'W) to evaluate the efficacy of a new insect growth regulator material (dicyclanil) in the prevention of myiosis caused by screwworm (Cochliomyia hominivorax) larvae. Forty bull calves (Bos indicus x Bos taurus), 5 to 6 months old were allocated by ranked paris to a control (G.1) and a treated group (G.2) of the same number of animals. On day 0, all experimental animals were surgically castrated and calves of group 2 were treated topically with 20 ml as a single dose of a ready-to-use formulation containing 5% w/v dicyclanil. Inspections for screwworm larva infestation were carried out on days 4, 8, 12, 16, 19, 23 and 25 after castration. The incidence of egg masses on the scrotal wounds evidenced a similar challenge for all groups (P > 0.05 chi(2)). However, 16 animals of G.1 (80%) and only one of G.2 (5%) developed active myiosis, respectively. This difference was significant (P=0.003 X(2)).

Babesial antibody dynamics after cattle immunisation with live vaccines, measured with an indirect immunofluorescence test.

The efficacy of vaccination of Argentinean cattle against babesiosis and anaplasmosis using live immunogens was tested to detect specific antibodies in samples obtained about 60 days after vaccination. Under these conditions a higher than expected proportion of cattle failed to show antibodies against Babesia bigemina. Therefore, a study was designed to evaluate if this failure was due to insensitivity of the routine test to detect antibodies to B. bigemina or to lack of infectivity of the live vaccine. Four groups (G) of cattle were each inoculated subcutaneously with 10 million Babesia bovis (vaccinal strain R1A), 10 million B. bigemina (vaccinal strain S1A) and 10 million Anaplasma centrale (strain M1). G1 and G2 consisted of ten Angus bulls 20-24 months old and ten Angus bulls 15-18 months old, respectively; G3 and G4 consisted of ten and 16 Holstein 1-month-old male calves, respectively. Blood samples were obtained on days 0, 10, 20, 30, 40, 50 and 60 after vaccination and the sera were analysed with an indirect immunofluorescent (IFA) test to detect antibodies to B. bovis (baseline dilution for a positive result 1:60) and B. bigemina (baseline dilution 1:120). Positive IFA titres were considered as evidence of the infectivity of the Babesia vaccinal strains contained in the vaccine. All Angus bulls were found positive to antibodies against both Babesia species, by day 20 (B. bovis) and day 30 (B. bigemina), whereas 10-25% of Holstein calves were negative throughout. The partial lack of vaccine infectivity in the calves was considered to be a consequence of innate resistance of young calves to Babesia. Antibody titres to B. bovis and B. bigemina declined by day 60 after vaccination. However, all cattle that were positive to B. bovis antibodies on day 50 were still positive to the IFA test 10 days later while 10%, 30% and 12% of cattle of G1, G2 and G3 that were positive to B. bigemina antibodies on day 50 after vaccination were found negative to the IFA test on day 60. In future, samples taken on days 40-50 will be used for detection of B. bigemina antibodies in vaccinated cattle, on day 60 for A. centrale and on either occasion for B. bovis. The reaction to the inoculation of B. bigemina S1A strain appears to lag behind the reaction to B. bovis R1A strain. It is not certain if this is a normal reaction to this B. bigemina strain or the result of interaction with the B. bovis strain.