Mechanism of resistance to synthetic pyrethroids in buffalo flies in south-east Queensland.

Resistance to synthetic pyrethroids (SP) was first recorded in buffalo flies in Australia in 1980, associated with previous use of DDT and fenvalerate. By the 1990s, resistance was widespread. Resistance to SP in the related horn fly of the Americas is associated with kdr and super-kdr mutations in a gene encoding for a voltage-gated sodium channel. We describe 7-20-fold resistance to SP in buffalo flies from south-east Queensland, present evidence of flies that are heterozygous resistant at the kdr locus and show an increase in the frequency of the resistant allele 1 month after treatment of cattle with SP.

Histological changes to the skin of Merino sheep following deep dermal and subcutaneous injections of sodium lauryl sulfate.

OBJECTIVES: To characterise the changes caused to sheep skin by deep dermal and subcutaneous injections of sodium lauryl sulfate (SLS) and describe the subsequent healing process. PROCEDURE: On 6 sheep 20 small areas of skin were each given deep dermal and subcutaneous injections of 0.2 mL of 7% SLS. Biopsies were collected at intervals up to 28 days after treatment and the histological changes in each of the treated skin samples were assessed and graded. RESULTS: There was no evidence of alterations in behaviour, weight gain or appetite of the sheep following the injections. Initial swelling of the treated site subsided by day 14, leaving a firm, slightly raised crust. At day 21, the treated area was depressed and covered by a scab, which sloughed completely by day 28. There was necrosis of the subcutis and deep dermis 2 min after treatment, followed by inflammation, fibroplasia, angiogenesis and tissue remodelling. CONCLUSION: Injection of SLS caused almost immediate local necrosis followed by eschar formation, sloughing and scarring of treated skin. Deep dermal and subcutaneous SLS is potentially useful as a chemical alternative to mulesing.

Rotation of treatments between spinosad and amitraz for the control of Rhipicephalus (Boophilus) microplus populations with amitraz resistance.

A farmlet study was conducted over 4 years in which three treatments were applied to six groups of Holstein dairy calves. Calves in each group were infested with equal numbers of N-strain (susceptible) and Ultimo strain (amitraz and synthetic pyrethroid resistant) tick larvae to establish self-sustaining populations with an initial, measurable level of resistance to amitraz. Standard counts of all ticks between 4.5 and 8.0mm diameter on one side of each animal were made each week and treatment was applied when tick numbers exceeded a threshold of 25 engorged adults per side. The three treatments were: 1, spinosad spray whenever tick numbers exceeded the threshold; 2, amitraz spray whenever tick numbers exceeded the threshold; 3, spinosad whenever tick numbers exceeded the threshold for the first 2 months, then amitraz for 2 months, with alternation every subsequent 2 months. Engorged adult female ticks were collected from each treatment group on 10 or 11 occasions during the study and tested using the larval packet test bioassay (LPT) for acaricide resistance. Spinosad 250ppm provided effective control of amitraz-resistant tick populations in the field, using a similar number of treatments as in the amitraz and rotation groups. The initial infestations of all of the groups resulted in the establishment of populations with in vitro evidence of resistance to amitraz using the LPT. Treatment with spinosad or with a rotation between spinosad and amitraz every 2 months resulted in reduced levels of resistance to amitraz according to the LPT. The animals treated with amitraz alone showed increasing resistance to amitraz according to the LPT each summer and autumn with a return to full or almost full susceptibility to amitraz in early spring in all years. This pattern suggests a relative lack of fitness of amitraz-resistant ticks that might be exploited by using an acaricide rotation strategy.

Effectiveness of a non-surgical alternative to the Mules operation in sheep.

OBJECTIVE: To measure changes to the perineal bare area, local tissue reaction and healing responses of young sheep, following intradermal administration of cetrimide and polyvinylpyrrolidone (PVP), with and without ethanol, to the breech and tail. METHOD: A needle-less injector was used to deposit formulations containing 40 g/L cetrimide and 30 g/L PVP (group 2) or 20 g/L cetrimide, 30 g/L PVP and 15 g/L ethanol (group 3), within the dermis of the tail and the region surrounding the perineal bare breech area of groups (N = 8) of Merino weaner sheep. The dimensions of the perineal bare area (length, width and diagonal distances left and right) and tail width were recorded before and at intervals after treatment for 60 days. Observations of swelling and bruising and scab formation at the treatment sites were recorded for up to 35 days after treatment. Rectal temperatures were monitored for up to 35 days after treatment and bodyweight for up to 60 days after treatment. An untreated control group (group 1) was included. RESULTS: Comparison of day -3 and day 35 measurement data showed that both treated groups had significantly (P < 0.05) wider breech bare areas compared to the untreated controls and that group 2 sheep had significantly (P < 0.05) longer breech bare areas compared to group 3 sheep or to the untreated controls, which were not significantly different. At this time scabs were still firmly in place on many treated sheep. At day 35 there was no increase in tail bare area caused by either treatment. By day 60 there was no significant difference between the treated and control groups in either the breech or tail regions indicating that the changes present at day 35, were not permanent. Mean weight gain in the groups throughout the 60-day interval was unaffected by treatment. Intradermal treatment was associated with a significant elevation in body temperature. This effect lasted for 3 days and was associated with signs of discomfort and depressed appearance in at least some of the treated sheep. Bruising was mild to severe in all treated sheep within two days of treatment but was not evident in any sheep by day 21. Mild to moderate swelling was also associated with treatment but was not uniform across sheep in the groups. The tail of one sheep was severely swollen for several days. Swelling remained obvious in most treated sheep until day 14 but was not present at day 21. CONCLUSION: Under the conditions of this study intradermal injection of cetrimide had no permanent effect on bare area measurements on the breech or the amount of wool-bearing skin on the tail. It also caused signs of discomfort and pain that raise welfare concerns.

Residues of spinosad in the tissues of sheep after aerosol treatment of blowfly myiasis.

OBJECTIVE: To measure the residues of spinosad and chlorhexidine in the tissues of sheep after treatment of blowfly strike. PROCEDURE: Fourteen sheep with natural myiasis and 12 with artificial infestations of Lucilia cuprina larvae had the wool removed over their infestations and were treated with an aerosol wound dressing containing spinosad and chlorhexidine. Sheep were killed up to 14 days after treatment and residues of the chemicals measured in tissues. RESULTS: Chlorhexidine was not detected in any tissue. Residues of spinosad were highest in fat, lowest in muscle and intermediate in liver and kidney. The highest residue detected was 0.2 mg/kg spinosad in perirenal fat 7 days after generous treatment of a sheep with a large fly strike. Residues of spinosad in fat peaked 3 to 7 days after treatment and 1 to 3 days after treatment in liver and kidney. CONCLUSION: These studies present a realistic worst-case in struck sheep and at the highest dose studied, equivalent to 5.8 mg spinosad per kg body weight, the maximum residue detected of 0.2 mg/kg in peri-renal fat was 20% of the Australian maximum residue limit. Muscle, liver and kidney residues of spinosad were also below the Australian maximum residue limits at all times.

Residues of zeta-cypermethrin in bovine tissues and milk following pour-on and spray application.

The depletion of zeta-cypermethrin residues in bovine tissues and milk was studied. Beef cattle were treated three times at 3-week intervals with 1 ml 10 kg-1 body weight of a 25 g litre-1 or 50 g litre-1 pour-on formulation (2.5 and 5.0 mg zeta-cypermethrin kg-1 body weight) or 100 mg kg-1 spray to simulate a likely worst-case treatment regime. Friesian and Jersey dairy cows were treated once with 2.5 mg zeta-cypermethrin kg-1 in a pour-on formulation. Muscle, liver and kidney residue concentrations were generally less than the limit of detection (LOD = 0.01 mg kg-1). Residues in renal-fat and back-fat samples from animals treated with 2.5 mg kg-1 all exceeded the limit of quantitation (LOQ = 0.05 mg kg-1), peaking at 10 days after treatment. Only two of five kidney fat samples were above the LOQ after 34 days, but none of the back-fat samples exceeded the LOQ at 28 days after treatment. Following spray treatments, fat residues were detectable in some animals but were below the LOQ at all sampling intervals. Zeta-cypermethrin was quantifiable (LOQ = 0.01 mg kg-1) in only one whole-milk sample from the Friesian cows (0.015 mg kg-1, 2 days after treatment). In whole milk from Jersey cows, the mean concentration of zeta-cypermethrin peaked 1 day after treatment, at 0.015 mg kg-1, and the highest individual sample concentration was 0.025 mg kg-1 at 3 days after treatment. Residues in milk were not quantifiable beginning 4 days after treatment. The mean concentrations of zeta-cypermethrin in milk fat from Friesian and Jersey cows peaked two days after treatment at 0.197 mg kg-1 and 0.377 mg kg-1, respectively, and the highest individual sample concentrations were 2 days after treatment at 0.47 mg kg-1 and 0.98 mg kg-1, respectively.

The binding of closantel to ovine serum albumin, and homogenate fractions of Haemonchus contortus.

Closantel binds to the serum proteins of the host and affects blood sucking parasites when they ingest the blood of treated hosts. Closantel binds specifically to ovine serum albumin (K(a) of 9. 3x10(6)M(-1)) at site I, the warfarin/phenylbutazone binding site of albumin Closantel also binds to invertebrate haemocyanin and haemolymph. The strongest binding of closantel in homogenates of H. contortus is found in fractions containing soluble proteins. This binding is of low affinity and, because the site itself is not fully denaturable, it may not be proteinaceous. There is no detectable difference in binding affinity between homogenate fractions from closantel susceptible and resistant isolates of adult or larval worms suggesting that closantel resistance is not due to changes in the closantel receptor or carrier.

Therapeutic efficacy of zeta-cypermethrin pour-on for the treatment of biting and sucking lice in cattle under field conditions.

OBJECTIVE: To assess the efficacy of zeta-cypermethrin pour-on to control cattle lice. DESIGN: Five field trials in south-eastern Australia. PROCEDURE: Zeta-cypermethrin pour-on, deltamethrin pour-on and pour-on vehicle were applied to groups of 10 cattle. Lice were counted before treatment and 14, 28, 42 and 56 days after treatment. RESULTS: Zeta-cypermethrin pour-on given at 2.5 mg/kg was equivalent to, or marginally more effective than a deltamethrin pour-on at 0.75 mg/kg. It eliminated B bovis and H eurysternus and gave good control of L vituli and S capillatus. Zeta-cypermethrin at 1 mg/kg gave good control of B bovis and H eurysternus but was not satisfactory against L vituli and S capillatus. CONCLUSION: Zeta-cypermethrin pour-on, given at 2.5 mg/kg, is an effective treatment for cattle lice control. Zeta-cypermethrin, and other synthetic pyrethroid pour-ons, are the treatment of choice to control B bovis.

Efficacy of zeta-cypermethrin as pour-on or spray formulations for the control of buffalo fly (Haematobia irritans exigua) in cattle.

OBJECTIVE: To determine the efficacy of zeta-cypermethrin in controlling buffalo fly (Haematobia irritans exigua). DESIGN: Five field trials in northern and central Queensland. PROCEDURE: Zeta-cypermethrin pour-on at 2.5 mg/kg, spray at 62.5 ppm, deltamethrin pour-on and pour-on vehicle were applied to groups of 20 cattle. Buffalo fly counts were conducted three times before treatment and 3, 7, 14, 21, 28 and 35 days after treatment. RESULTS: In central Queensland where synthetic pyrethroid resistance in buffalo fly populations was rare, 2.5 mg/kg of zeta-cypermethrin pour-on gave good control of buffalo fly for 4 weeks and was better than a deltamethrin product. A zeta-cypermethrin spray used at 62.5 ppm gave 14 days control. In far-north Queensland where resistance to synthetic pyrethroids and heavy rain was common, the maximum period of efficacy of zeta-cypermethrin pour-on was reduced to 2 weeks. CONCLUSION: In areas where there is low resistance to synthetic pyrethroids among buffalo flies, zeta-cypermethrin pour-on can be expected to give good control for 4 weeks.

Comparative in vitro effects of closantel and selected beta-ketoamide anthelmintics on a gastrointestinal nematode and vertebrate liver cells.

PNU-87407 and PNU-88509, beta-ketoamide anthelmintics that are structurally related to each other and to the salicylanilide anthelmintic closantel, exhibit different anthelmintic spectra and apparent toxicity in mammals. The basis for this differential pharmacology was examined in experiments that measured motility and adenosine triphosphate (ATP) levels in larval and adult stages of the gastrointestinal nematode, Haemonchus contortus, and in a vertebrate liver cell line and mitochondria. PNU-87407 and PNU-88509 both exhibited functional cross-resistance with closantel in larval migration assays using closantel-resistant and -sensitive isolates of H. contortus. Each compound reduced motility and ATP levels in cultured adult H. contortus in a concentration- and time-dependent manner; however, motility was reduced more rapidly by PNU-88509, and ATP levels were reduced by lower concentrations of closantel than the beta-ketoamides. Tension recordings from segments of adult H. contortus showed that PNU-88509 induces spastic paralysis, while PNU-87407 and closantel induce flaccid paralysis of the somatic musculature. Marked differences in the actions of these compounds were also observed in the mammalian preparations. In Chang liver cells, ATP levels were reduced after 3 h exposures to > or = 0.25 microM PNU-87407, > or = 1 microM closantel or > or = 10 microM PNU-88509. Reductions in ATP caused by PNU-88509 were completely reversible, while the effects of closantel and PNU-87407 were irreversible. PNU-87407, closantel and PNU-88509 uncoupled oxidative phosphorylation in isolated rat liver mitochondria, inhibiting the respiratory control index (with glutamate or succinate as substrate) by 50% at concentrations of 0.14, 0.9 and 7.6 microM, respectively.

Occurrence of cutaneous infections of Myxobolus episquamalis (Myxozoa:Myxobolidae) in sea mullet, Mugil cephalus L, in Australia.

OBJECTIVE: To report the occurrence of Myxobolus episquamalis in sea mullet, Mugil cephalus L, caught in estuaries in eastern and western Australia. DESIGN: A prospective study of commercial catches of mullet in the Clarence River of NSW and individual cases from other areas. RESULTS: The organism caused pale, white to pink, raised lesions on the scales and fins of sea mullet. Occurrence of infection was highest in spring and in a marine (down-river) environment compared to a brackish environment. Up to 6% of fish were affected in commercial catches. CONCLUSION: The infection is widespread in Australian mullet, but rarely causes significant economic loss.

The effects of closantel treatment on the ultrastructure of Haemonchus contortus.

H. contortus were recovered from sheep 0-14 h after intramuscular treatment with closantel. Ultrastructural examination revealed that mitochondria were more electron dense and contained swollen cristae compared with untreated controls. Following treatment, the basal channels in the intestine became prominent and there was vesicle formation in all organs examined. In contrast, closantel-resistant H. contortus appeared normal after drug treatment. It is likely that closantel affects membrane associated processes responsible for fluid and ion homeostasis as well as mitochondrial function. Untreated H. contortus were maintained in balanced salt solution for 12 h which caused lesions indicative of fluid imbalance, but at 23 h there were serious structural abnormalities.

Gossypol toxicity in preruminant calves.

Twenty-four of 57 calves fed a diet containing 33% cotton seed meal (CSM) died between 7 and 15 weeks of age. Initial deaths were not accompanied by premonitory signs, but after CSM withdrawal most calves developed rough coats, anorexia, weakness, ascites and subcutaneous oedema. Those that died had large volumes of serous fluid in the body cavities, hard livers of 'nutmeg' appearance, and pulmonary congestion. Histopathologically the livers showed periacinar necrosis in acute cases and periacinar fibrosis in chronic cases. Lungs from several calves had oedema, haemosiderosis and fibrosis in some pulmonary vessels. Atrophy of myocardial fibres was present in most cases. The concentration of free gossypol in the diet was 100 to 220 mg/kg. Ante-mortem and post-mortem findings supported a diagnosis of gossypol poisoning. The deaths continued for 4 weeks after withdrawal of CSM from the diet.

Colorectal blood supply in dogs.

To determine blood supply in the area, dye or radioopaque contrast material was injected into the named arteries supplying the terminal colon and rectum in 10 dogs. The cranial rectal artery appeared to supply most of the blood to the terminal colon and rectum. The middle and caudal rectal arteries supplied variable and relatively insignificant amounts. The intrapelvic rectum had a less adequate blood supply than did the terminal colon or proximal rectum. A ventral midline laparotomy and pubic osteotomy were performed in an additional 11 dogs to provide access to the terminal colon and rectum. When the cranial rectal artery was ligated and the colorectal junction was transected and anastomosed, the intrapelvic rectum developed marked congestion, edema, and discoloration. Rectal fluorescence, after IV administration of fluorescein, was either poor or absent. Histologically, partial- to full-thickness mucosal necrosis was evident in most tissue specimens taken from the rectum, and muscle necrosis was evident in some. These findings suggest that, in dogs, the cranial rectal artery should be preserved if at all possible and, if the cranial rectal artery is ligated, most of the intrapelvic rectum should be resected to ensure adequate blood supply to the anastomosis.

Kinetics of expulsion of Haemonchus contortus from sheep and jirds after treatment with closantel.

Experiments were conducted in sheep after intramuscular treatment with closantel and in jirds after oral treatment with closantel to determine when expulsion of established H. contortus commences. Expulsion starts at about 8 h in sheep and coincides with the onset of reduced motility in worms recovered from the abomasum. In jirds, expulsion starts by 2 h after treatment. Experiments also conducted in jirds showed that infective larvae are first killed by circulating closantel 3 days after infection, when blood feeding starts, and that by 8 days 80% of larvae are lost.

An in vitro assay utilising parasitic larval Haemonchus contortus to detect resistance to closantel and other anthelmintics.

A new assay for detecting resistance to anthelmintics in vitro is described. The assay uses a simple culture system in which the ability of anthelmintics to kill or inhibit the migration of parasitic third and fourth stage Haemonchus contortus larvae through a 50 microns aperture mesh is assessed. The assay detects 2-10-fold resistance to closantel. Resistance to benzimidazoles, levamisole and ivermectin can also be detected.

Meningoencephalitis and other conditions associated with Histophilus ovis infection in sheep.

Histophilus ovis was isolated from 29 sheep in 20 flocks and 2 artificial insemination (AI) centres in southern New South Wales from 1984 to 1990. The clinical and pathological findings were consistent with previous reports and included polyarthritis (7 flocks), epididymo-orchitis (5), meningoencephalitis (3), pneumonia (3), septicaemia (2), mastitis (1) and metritis (1). Six sheep had meningoencephalitis, a syndrome not previously associated with H ovis infection in sheep, which was similar pathologically to thromboembolic meningoencephalitis in cattle, caused by the related organism, Haemophilus somnus. H ovis was isolated from the semen of 12-month-old rams in a flock that had polyarthritis due to H ovis, in 4-month-old ram lambs and from the uterus of a ewe in a flock that had sporadic cases of H ovis septicaemia.

Replication of bovine viral diarrhoea virus in the bovine reproductive tract and excretion of virus in semen during acute and chronic infections.

Five mature bulls were studied during an acute transient infection with bovine viral diarrhoea virus (BVDV). The bulls had been infected experimentally by the intranasal instillation of blood and serum from a cow which was a persistent carrier of the virus. Infection was confirmed by the demonstration of a low titred viraemia in four of the five animals and by the seroconversion of all five. Semen samples were collected from each bull on four occasions between seven and 14 days after infection. The virus was isolated from the semen of three of the five bulls and from nine of 12 batches of semen from them. In contrast to other studies of the infection of semen, BVDV was isolated with similar efficiency from raw, unprocessed semen and from diluted, extended semen. The titres of virus in the semen ranged from 5 to 75 TCID50/ml. The infection did not appear to affect the quality of the semen. Shedding of virus continued after the end of the period of viraemia and appeared to be a consequence of the replication of the virus in the reproductive tract and its subsequent excretion in the seminal fluid. Virological studies of the reproductive tracts of these bulls suggested that the most productive sites of virus replication were the seminal vesicles and the prostate gland. Concurrent studies in a persistently infected bull supported these findings.