Investigations into the Prevention of Neonatal Ancylostoma caninum Infections in Puppies by Application of Moxidectin to the Bitch

The aim of this investigation was to examine whether reactivated Ancylostoma caninum larvae can be eliminated by the administration of moxidectin to pregnant bitches. Four pregnant bitches infected experimentally with 20 000 third-stage larvae of A. caninum were treated subcutaneously with 1 mg moxidectin/kg body weight on day 55 of the pregnancy (5–8 days before parturition). Another four experimentally infected pregnant bitches served as controls. The single moxidectin treatment completely prevented lactogenic infections in the puppies. Neither intestinal stages nor somatic larvae could be found. The administration of moxidectin caused no local or systemic side-effects in the bitches. All 22 puppies of the treated bitches were born healthy and remained so during the whole trial period. Beginning during the third week after birth, all 20 puppies of the untreated bitches developed a severe microcytic, hypochromic anaemia and they revealed a total of 8649 intestinal stages of A. caninum after autopsy.     

Investigations into the prevention of neonatal Ancylostoma caninum infections in puppies by application of imidacloprid 10% plus moxidectin 2.5% topical solution to the pregnant dog

The aim of the investigation was to examine whether a single topical administration of a combination of imidacloprid and moxidectin to pregnant dogs could prevent neonatal infections with reactivated Ancylostoma caninum larvae. Three pregnant beagles, infected with A. caninum, were treated topically with the combination on day 56 of pregnancy. Three further dogs served as untreated controls. Treatment appeared to prevent neonatal infections in the puppies completely. Neither intestinal stages nor somatic larvae were found in two examined puppies per litter. All puppies and dams of the treatment group remained coproscopically negative. No side-effects in dams or puppies were observed. Two of three untreated dams showed a patent infection after parturition. Necropsy of two puppies of each negative control litter revealed seven intestinal and five somatic A. caninum stages in total. One litter of the untreated dams showed a patent infection 33 days after parturition. In the other two litters, no representative sample sizes could be collected.     

Investigations into the prevention of prenatal and lactogenic Toxocara canis infections in puppies by application of moxidectin to the pregnant dog

Aim of the investigation was to examine whether two administrations of moxidectin to pregnant dogs could prevent pre-natal and lactogenic infections of puppies with reactivated Toxocara canis larvae. Four pregnant beagles, infected experimentally with 20 000 embryonated eggs of T. canis, were treated subcutaneously with 1 mg moxidectin per kg body weight on days 40 and 55 of pregnancy (5-13 days before parturition). One further dam and its puppies served as untreated control. Two applications of moxidectin completely prevented pre-natal and lactogenic infections in the puppies. Neither intestinal stages nor somatic larvae were found in the dams or their corresponding puppies. All puppies and dams of the treatment group remained coproscopically negative until 42 days after parturition. The administration of moxidectin did not show any side effects in the dams. None of the puppies of the treated dams showed any pathological abnormalities. In the untreated dam one adult and 26 somatic larvae of T. canis were detected at necropsy. All puppies of the untreated dam showed a patent T. canis infection from day 28 post-natum (p.n.); 296 pre-adult and adult stages of T. canis were spontaneously eliminated and 51 intestinal stages and five somatic larvae of T. canis were recovered at necropsy. In contrast to the puppies of the treated dams all negative control puppies showed blood eosinophilia after parturition and elevated liver enzyme levels.     

The effect of ivermectin on reactivated somatic larva of Ancylostoma caninum Ercolani 1859 (Ancylostomidae) in the pregnant dog

It was evaluated by means of a controlled test to what extent reactivated larvae of Ancylostoma caninum were influenced by single or double treatment with ivermectin. The investigations were done with a total of ten experimentally infected bitches. The single treatment of dams two to ten days ante partum with 0.5 mg or 1 mg ivermectin per kg bodyweight reduced the wormburden of their puppies on an average by 96.6% and 98.5%, respectively. The double treatment with 0.5 mg/kg bodyweight ante and post partum each with an interval of ten days completely prevented lactogenic infections with Ancylostoma caninum. Following a single application of 0.5 mg ivermectin per kg bodyweight the excretion of larvae was markedly delayed and remained on a low level. In addition the infections of the puppies of single treated dams became patent on an average 9.5 and 14.5 days later than the controls. In the bitches no side-effects, neither local nor general, were seen after the treatment with ivermectin. All forty puppies of the treated bitches were born without visible damages.     

Parasitologic, clinical, hematologic and serologic findings in puppies after lactogenic infection with Ancylostoma caninum ERCOLANI 1859 (Ancylostomidae)]

The correlation between intensity of Ancylostoma caninum infections in bitches and the intensity of lactogenic infections and clinical signs of their puppies was investigated. On average, 825, 1,867 or 2,125 specimens were observed in litters of two bitches inoculated with 5,000, 10,000 or 20,000 third stage larvae (LIII) respectively, at the day of conception. Adverse effects of the infection on the growth and behaviour of the puppies were observed after onset of their second week of life: 11/27 puppies died during the fourth week. The body weight of puppies surviving 28 days was up to 750 g less than that of uninfected controls. Eosinophilia, erythroblastosis and microcytic, hypochromic anemia developed in all puppies during their first four weeks. The IFA test (LIII antigen) was positive for only 5/18 heavily infected puppies after uptake of colostrum.     

Biology, pathogenicity, diagnosis and control of Ancylostoma caninum]

Infections with Ancylostoma caninum are transmitted orally or percutaneously. The transmission of infectious stages with the milk of particular importance for the distribution of the species. It occurs during the dissemination of larvae that follows every infection as well as after reactivation of resting somatic larvae in the bitch at the end of the pregnancy. The galactogenic transmission of larvae occurs even when, due to existing immunity, no patent infections develop in the bitch. Immunity does not or only to a low extent influence impatient infections or the migration of reactivated somatic larvae. It also allows a limited reestablishment of a deposit of larvae in the bitch. Following percutaneous infection dermatitis occurs in the area of larval penetration and the lung is affected by migrating larvae. Intestinal stages of Ancylostoma caninum damage the host by ingestion of the mucosa of the small intestine and withdrawal of blood. Main symptoms of ancylostomiasis are a mucous haemorrhagic diarrhoea and anaemia, that become visible 8 to 10 days post infection. The examination for impatient infections with Ancylostoma caninum can be done by immunofluorescence and ELISA. With both methods antibodies against third stage larvae can be detected from the first or second week post infection onward. Patent infections with Ancylostoma caninum can easily be detected by faecal examination for the presence of the characteristic oval, thin-walled eggs containing few blastomeres. Galactogenic infections with Ancylostoma caninum can be prevented or reduced by a regular treatment of the bitch with albendazole, fenbendazole or oxfendazole during the activation of larvae in the last third of the pregnancy or by repeated treatment with ivermectin shortly before and after birth. To prevent patent infections, galactogenic infected puppies have to be treated early and repeatedly.     

Six-month prophylactic efficacy of moxidectin sustained release (SR) injectable for dogs against experimental heartworm infection in growing puppies

The purpose of this study was to assess the efficacy of moxidectin sustained release injectable for dogs (moxidectin SR, Fort Dodge Animal Health) in protecting growing puppies from experimental infection with the heartworm, Dirofilaria immitis, six months after treatment. The study involved 27 puppies, approximately 12 weeks of age at the beginning of the study, with nine puppies in each of three size classes. The small breed class included eight Pekingese and one purpose-bred small breed mongrel; the medium breed class included nine purpose-bred mongrels, and the large breed class included nine puppies with an anticipated adult weight >or=30-35 kg. Both genders were included with no attempt made to have equal numbers of male and female puppies. Puppies were blocked by weight within each size class and randomly assigned to three treatment groups of nine dogs. On Day 0, pups in two groups were injected subcutaneously with moxidectin SR, dosed to deliver 0.17 mg moxidectin/kg b.w. The third group was injected with sterile saline. Personnel making observations were blinded to the treatment status of the animals. Following treatment, puppies were observed for signs of adverse local and systemic reactions. Puppy weights and serum moxidectin levels were also monitored. On Day 180, puppies in all treatment groups were inoculated subcutaneously with 50 third-stage larvae of D. immitis. On Days 348 and 349, puppies were euthanatized and necropsied. Hearts and lungs were examined for adult heartworms. All animals in the saline control group were infected with an arithmetic mean of 39.22 adult heartworms each. Seventeen of 18 dogs in the moxidectin SR-treated groups were uninfected. One treated puppy was infected with a single adult heartworm. This infected individual was from the large breed size class and had the second highest percent increase in body weight. Based on arithmetic means, the heartworm recovery from all treated puppies represents a 99.86% reduction relative to the saline control. There were no adverse local or systemic reactions to treatment in any animal.     

Persistent efficacy of moxidectin canine sustained-release injectable against experimental infections of Ancylostoma caninum and Uncinaria stenocephala in dogs

The efficacy of moxidectin canine sustained-release injectable administered at fixed intervals before administration of an oral hookworm challenge was evaluated in 40 laboratory dogs. Groups of eight dogs were treated with the moxidectin sustained-release formulation by SC injection approximately 3, 4, 5, or 6 months before being given oral inoculations with 300 Ancylostoma caninum larvae on Day 0 and 400 Uncinaria stenocephala larvae one day later. Dogs were euthanized 21 days after parasite inoculations. Fecal samples and the intestinal contents collected at necropsy from each dog were examined for hookworms. Fecal egg count reductions based on geometric means relative to controls were 99.2% (3 months) to 81.2% (6 months). The reduction in A. caninum worm recoveries at 3, 4, 5, and 6 months based on geometric means were 94.7%, 90.3%, 82.0%, and 60.2%, respectively. The mean reductions in U. stenocephala worm counts at these intervals were 94.6%, 85.3%, 71.6%, and 48.2%,respectively.     

Fenbendazole treatment of pregnant bitches to reduce prenatal and lactogenic infections of Toxocara canis and Ancylostoma caninum in pups

A granulated formulation of fenbendazole was tested in a total of 23 treated and control, pregnant, parasite-free Beagle bitches experimentally infected with Toxocara canis and Ancylostoma caninum. The drug was administered to each treated bitch once daily in canned dog food at a dosage of 50 mg/kg body weight. Each of 2 treatment regimens tested was initiated on the 40th day of pregnancy. One regimen involved daily treatment continuing through the 14th postpartum day, and it resulted in 89% fewer ascarids and 99% fewer hookworms in pups born to medicated bitches, as compared with pups born to unmedicated controls. The other regimen of treatment, which was stopped on the day of parturition, was less effective in reducing ascarid and hookworm burdens (64% and 88% reductions, respectively). Three to 5 bitches from each of the treatment and control groups were allowed to whelp a 2nd litter without further treatment or further exposure to parasite infections. Hookworm burdens in 2nd-litter pups born of bitches that had initially received fenbendazole through the 14th postpartum day were significantly lower (P < 0.01; 85% reduction), when compared with the 2nd-litter control pups. All other parasite burdens were not significantly different. It was concluded that granulated fenbendazole is effective in reducing burdens of Ancylostoma caninum and Toxocara canis in newborn pups when the bitch is treated during the last third of pregnancy, especially when treatment (50 mg/kg/day) extends from the 40th day of pregnancy through the 14th postpartum day.     

Toxocara canis infection induces antigen-specific IL-10 and IFNgamma production in pregnant dogs and their puppies

Toxocara canis (T. canis) is originally a parasite of canine bitches and their pups. The pathogenicity of T. canis infection is enhanced during pregnancy and puppyhood. The aim of this study was to investigate if modification of IFNgamma and IL-10 secretion occurs during infection in pregnant dogs and puppies. Analysis of cytokines secreted could let us hypothesize a role for IL-10 and/or IFNgamma in T. canis infection. We tested T. canis-specific production of IFNgamma and IL-10 by lymphocytes of pregnant dogs and their puppies after in vitro re-exposure to purified excretory/secretory antigen (ESAg) from T. canis. Blood mononuclear cells (BMC) isolated from pregnant dogs and their puppies were cultured in the presence of ESAg. Cultures' supernatants were tested for cytokine levels by ELISA. Results obtained showed that IL-10 concentrations increased during pregnancy in infected animals and in the meantime IFNgamma production decreased. In puppyhood, we observed that, IL-10 concentration decreased with the age of puppies mainly in infected animals while IFNgamma increased. In conclusion, our data suggests that BMC of infected dogs have a particular modification of IL-10 and IFNgamma synthesis. These data could be the basis to design immunotherapeutic approaches.     

Transmission of Babesia spp by the cattle tick (Boophilus microplus) to cattle treated with injectable or pour-on formulations of ivermectin and moxidectin

OBJECTIVE: To assess the efficacy of ivermectin and moxidectin to prevent transmission of Babesia bovis and Babesia bigemina by Boophilus microplus to cattle under conditions of relatively intense experimental challenge. DESIGN: Naive Bos taurus calves were treated with either pour-on or injectable formulations of either ivermectin or moxidectin and then exposed to larvae of B microplus infected with B bovis or larvae or adults of B microplus infected with B bigemina. One calf was used for each combination of haemoparasite, B microplus life stage, drug and application route. PROCEDURE: Groups of calves were treated with the test drugs in either pour-on or injectable formulation and then infested with B microplus larvae infected with B bovis or B bigemina. B bigemina infected adult male ticks grown on an untreated calf were later transferred to a fourth group of animals. Infections were monitored via peripheral blood smears to determine haemoparasite transmission. RESULTS: Cattle treated with either pour-on or injectable formulations of ivermectin and moxidectin became infected with B bovis after infestation with infected larvae. Similarly, larvae infected with B bigemina survived to the nymphal stage to transmit the haemoparasite to animals treated with each drug preparation. Cattle treated with pour-on formulations of ivermectin and moxidectin then infested with adult male ticks infected with B bigemina did not become infected with B bigemina whereas those treated with the injectable formulations of ivermectin and moxidectin did show a parasitaemia. CONCLUSIONS: Injectable or pour-on formulations of ivermectin and moxidectin do not prevent transmission of Babesia to cattle by B microplus. Use of these drugs can therefore not be recommended as a primary means of protecting susceptible cattle from the risk of Babesia infection.     

Clinical Evaluation of the Use of Aglepristone Associated with Oxytocin to Induce Parturition in Bitch

To evaluate the efficacy and safety of aglepristone 15 mg kg⁻¹ for induction of parturition in bitches, 22 pregnant beagle bitches were injected subcutaneously on day 60 post-estimated LH surge, and again 24 h later with aglepristone and subsequently were given 0.15 IU kg⁻¹ oxytocin at hourly intervals until delivery of the last puppy. Six pregnant beagle bitches were used as a non-treated control group. In the control group, parturition occurred at 63.2 ± 0.5 days, 29 pups were born and the average expulsion time per puppy was 1.0 ± 0.6 h. In the treated group, parturition was obtained on average 29.7 ± 5.6 h after aglepristone administration, 121 pups were born and average expulsion time per pup was 1.1 ± 0.4 h. The percentage of live puppies, 7 weeks after birth, was 86.1% (25/29) and 86.8% (105/121) for the control and treated groups, respectively. No significant difference was observed between the control and treated groups for the average expulsion time per live puppy and for the percentage of live puppies at birth, 48 h, 7 days or 7 weeks after birth (p > 0.05). This study confirms previous results and demonstrates that the combination of aglépristone and oxytocin can be safely and reliably used to induce parturition in beagle bitches, at 60 days post-estimated LH surge.     

Safety evaluation of moxidectin sustained-release injectable in 10-week-old puppies

A study was conducted to evaluate the safety of a commercial formulation of moxidectin sustained-release injectable for dogs (ProHeart 6, Fort Dodge Animal Health) administered as a single subcutaneous dose to 10-week-old puppies. Twelve male and 12 female purpose-bred beagles 10 weeks of age were blocked by weight within gender and randomly allocated to three treatment groups. Puppies in two groups were treated with moxidectin sustained-release injectable for dogs at three or five times the labeled dose rate of 0.17 mg moxidectin/kg. The third group was treated with saline solution as controls. Physical and neurologic status, hematologic parameters, clinical chemistries, urine samples, body weight, and food consumption were evaluated before and up to 12 weeks after treatment. When compared to controls, mild depression of erythropoiesis, characterized by reduced hemoglobin, reticulocytes, erythrocytes, and hematocrit, was noted in puppies treated with five times the label dose of moxidectin sustained-release injectable. Values for these parameters remained within normal ranges and increased during the study, but at a reduced rate relative to saline-treated controls. Other parameters evaluated remained within normal limits for all treatment groups. Based on results of this study, the no observed adverse effect level for moxidectin sustained-release injectable (ProHeart 6) treatment in 10-week-old puppies was determined to be three times the recommended rate.     

Colonization of neonatal puppies by staphylococci.

The composition of the normal staphylococcal flora of bitches and their litters held in a breeding unit was studied. The animals were sampled at a number of sites using moistened swabs. Six bitches were sampled daily, for 10 days, before whelping and then, together with four puppies per litter, at whelping (day 11) and at 1 and 7 days thereafter. Staphylococcus intermedius formed the predominant staphylococcal isolate. S. intermedius counts at the oral and nasal sites on the bitches did not change markedly before whelping and remained low (< 6 cfu/swab). Significant rises in the oral counts on both the bitches (P < 0.05) and puppies (P < 0.001) were then observed after whelping (days 11-18). Abdominal counts on both the bitches and puppies also rose (P < 0.001) after whelping. S. intermedius counts at the vaginal vestibulum of the pregnant bitches were found to be higher than at any other site sampled and did not alter markedly until whelping when a decrease (P = 0.05; days 10-12) was observed. S. intermedius was not found at the anal site in any of the six bitches and only transiently colonized five of the puppies.     

Imidacloprid/moxidectin topical solution for the prevention of heartworm disease and the treatment and control of flea and intestinal nematodes of cats

Sixteen controlled laboratory studies, involving 420 kittens and cats, were conducted to evaluate the efficacy and safety of topically applied formulations of imidacloprid and moxidectin for the prevention of feline heartworm disease, treatment of flea infestations and treatment and control of intestinal nematodes. Unit-dose applicators and the dosing schedule used in these studies were designed to provide a minimum of 10mg imidacloprid and 1mg moxidectin/kg. Treatments were applied topically by parting the hair at the base of the skull and applying the solution on the skin. Imidacloprid treatment alone did not display activity against Dirofilaria immitis or intestinal nematodes and moxidectin treatment alone provided little or no activity against adult Ctenocephalides felis infestations. The formulation containing 10% imidacloprid and 1% moxidectin was 100% efficacious against the development of adult D. immitis infections when cats were treated 30 days after inoculation with third-stage larvae. A single treatment with this formulation also provided 88.4-100% control of adult C. felis for 35 days. Imidacloprid/moxidectin was 100% efficacious against adult Toxocara cati and 91.0-98.3% efficacious against immature adults and fourth-stage T. cati larvae. The formulation provided 98.8-100% efficacy against adult Ancylostoma and immature adults and third-stage A. tubaeforme larvae. Monthly topical application with 10% imidacloprid/1% moxidectin is convenient, efficacious and safe for the prevention of feline heartworm disease, treatment of flea infestation and for the treatment and control of intestinal nematode infections of cats.     

Efficacy of moxidectin against gastrointestinal nematodes of cattle.

Three groups of 11 naturally infected crossbred beef calves were injected subcutaneously with moxidectin 1 per cent injectable at 0.2 or 0.3 mg moxidectin/kg bodyweight or with the unmedicated vehicle. Nematode infections had been acquired during grazing from December to April. Based on the faecal egg counts and total worm counts of the control calves at necropsy (11 to 13 days after treatment) most of the calves had heavy parasitic burdens. Ostertagia ostertagi was predominant and the mean numbers of adults, developing fourth stage larvae (L4) and inhibited early L4 were 45,906, 10,061 and 68,918, respectively. Haemonchus placei and Trichostrongylus axei were also present in the abomasa. Three species of Cooperia, Oesophagostomum radiatum L4 and T colubriformis adults were found in the intestinal tract. Both dosages of moxidectin were equally effective (P < 0.05) against all the abomasal nematodes (99.9 to 100 per cent) and the intestinal tract nematodes (99.4 to 100 per cent). No adverse reactions to the moxidectin treatment were observed. Abomasal pathology characteristic of heavy O ostertagi infection was observed in the control calves, but not in the treated calves.     

Evaluating the accuracy of canine pregnancy diagnosis and litter size using real-time ultrasound

A total of 135 bitches suspected of being pregnant were examined with a real-time ultrasound equipment. 58 bitches were not pregnant. In pregnancy diagnosis ultrasound produced an overall accuracy of 99.3%, sensitivity 98.7%, and specificity 100.0%. Estimation of the litter size was fairly reliable in small litters of 1–3 puppies.     

Prenatal and transmammary infections of Toxocara canis in dogs: effect of benzimidazole-carbamate anthelmintics on various developmental stages of the parasite

Fenbendazole and albendazole, given at a dose rate of 150 mg/kg for 3 days, produced a 90 per cent reduction in the numbers of second stage larvae of Toxocara canis present in the tissues of dogs although no reduction in the number of larvae found in the brains of infected dogs occurred with this treatment. The results suggest that a course of 3 day therapy with these anthelmintics should prevent prenatal infections in puppies. However, if infection is acquired by bitches during late pregnancy or early lactation, the transmammary route of infection becomes important. Therefore, anthelmintic treatment of the bitch prior to pregnancy will not prevent transmission of infection to her puppies should the bitch acquire a new infection of T. canis during pregnancy or early lactation. Alternatively, infection with T. canis can be controlled through the treatment of neonatal puppies for migrating larvae of T. canis. Treatment of newborn puppies with fenbendazole, albendazole or oxfendazole at a dose of 100 mg/kg for 2–3 days produced a 91–99 per cent reduction in the number of adult parasites found. In addition, a single dose of fenbendazole, given at a dose rate of 40 mg/kg, eliminated 93–96 per cent of adult T. canis from the intestines of 4–5-week-old puppies. These latter treatments would need to be repeated to eliminate completely the infection from puppies.     

Effects of host maturity and prior exposure history on the production of Neospora caninum oocysts by dogs

To investigate whether dogs shed Neospora caninum oocysts more than once, five dogs with a previous history of shedding oocysts were fed infected bovine tissues. Two of three dogs shed oocysts when they were re-exposed 18-20 months after the first challenge; two other dogs re-exposed earlier, only 8 months after the primary exposure, did not produce oocysts. These results suggest that dogs may become refractory to shedding N. caninum oocysts for a period approximately between 8 and 18 months after a primary infection; however, this possibility requires statistical validation by testing of more dogs. The development of a high antibody titer did not ensure that a dog would completely resist shedding oocysts after consuming an infected meal. Oocyst production was also compared between puppies and adult dogs with primary infections. Twelve puppies (three from the present study and nine from a previous study) shed significantly more oocysts (mean: 166,400) compared with five adult dogs following primary exposure (mean: 2900), indicating that a dog's age can influence N. caninum oocyst production (P=0.02).     

Ivermectin treatment of naturally acquired and experimentally induced Strongyloides stercoralis infections in dogs.

Treatment of Strongyloides stercoralis infection was investigated in 2 dogs with naturally acquired, chronic-active infections, and in 3 dogs with corticosteroid-enhanced, experimentally induced hyperinfections. A single oral dose of ivermectin was given to naturally infected (200 micrograms/kg of body weight) and experimentally infected (800 micrograms/kg) dogs. Five dogs with experimental hyperinfections served as controls. Dogs with naturally acquired infections ceased to shed first-stage larvae in the feces 1 week after treatment, but 1 dog had recrudescence and required a second dose. Ivermectin was 100% effective in removing adult S stercoralis from the intestinal tract of the experimentally infected dogs, but it was not effective in removing third-stage larvae from parenteral sites. Ivermectin-treated dogs had few intestinal parasites of any stage, whereas at necropsy, 4 of 5 experimentally infected dogs not treated had massive infections (greater than 100,000 adults, greater than 92,000 larvae) in the intestinal tract, and 3 of 5 had larvae (greater than 2,500) in parenteral sites.