Safety of an extended-release injectable moxidectin suspension formulation (ProHeart® 12) in dogs.

BACKGROUND: The safety of ProHeart® 12 (PH 12; extended-release injectable suspension; 10% moxidectin in glyceryl tristearate microspheres) was evaluated in four studies using Beagle dogs and one study using ivermectin-sensitive Collies. The recommended dose is 0.5 mg/kg subcutaneously once yearly. METHODS: Study 1: safety margin was evaluated as 3 treatments of PH 12 (0× (control); 1× (recommended dose); 3× (3 times recommended dose) and 5× (5 times recommended dose) in 12 months via clinical observations, body weights, food consumption, injection site observations, physical examinations, moxidectin tissue assay, pharmacokinetics, and clinical and anatomic pathology. Study 2: safety in breeding-age males was demonstrated by semen testing at 14-day intervals from Day 7 to Day 91 post-treatment (0× or 3×). Study 3: reproductive safety in females was demonstrated by monitoring dams and litters following treatments (0× or 3×) administered during breeding, gestation, or lactation. Study 4: safety in dogs surgically implanted with adult heartworms was evaluated by clinical and laboratory monitoring following treatment with 0× or 3× administered 61 days post-implantation. Study 5: safety in ivermectin-sensitive dogs (120 µg/kg SC) was by clinical monitoring for 1 week after administering 1×, 3× or 5×. RESULTS: Study 1: slight swelling clinically detectable at some 3× and 5× injection sites was characterized microscopically as granulomatous inflammation, like tissue responses to medical implants, interpreted as non-adverse. Pharmacokinetics were dose-proportional and there was little or no systemic accumulation. Residual moxidectin mean (range) at 1× injection sites after 1 year was 16.0% (0.045-37.6%) of the administered mass. Studies 2 and 3: no effects were identified in reproductive indices (females) or semen quality characteristics (males). Study 4: PH 12 produced marked reductions in circulating microfilariae and lower numbers of adult heartworms, but no adverse clinical signs were identified. Study 5: there were no abnormal clinical signs at 1×, 3× or 5× overdoses of PH 12 in ivermectin-sensitive dogs. CONCLUSIONS: PH 12 has a > 5× safety margin in both normal and ivermectin-sensitive dogs, has no effects on canine reproduction, and is well tolerated in heartworm-positive dogs. The only treatment-related finding was non-adverse, granulomatous inflammation at the injection site.

Estimated direct economic costs associated with tick-borne diseases on cattle in Tanzania.

Tick-borne diseases, namely, anaplasmosis, babesiosis, cowdriosis and theileriosis, constrain cattle production and improvement in Tanzania, leading to considerable economic losses. A simple spreadsheet model was used to estimate the economic losses resulting from production losses, treatment and control costs associated with tick-borne diseases (TBD) in Tanzania. Model parameters included the national cattle population, reported TBD morbidity, fatality risk, and chemotherapy and control measures used. The total annual national loss due TBD was estimated to be 364 million USD, including an estimated mortality of 1.3 million cattle. Theileriosis accounted for 68% of the total loss, while anaplasmosis and babesiosis each accounted for 13% and cowdriosis accounted for 6% of the total loss. Costs associated with mortality, chemotherapy and acaricide application accounted for 49%, 21% and 14% of the total estimated annual TBD losses, respectively, infection and treatment method milk loss and weight loss accounted for 1%, 6% and 9% of the total annual loss, respectively. Despite the inadequacies of the data used, the results give evidence that tick-borne diseases inflict substantial economic losses on cattle production and resource use in Tanzania.

Determination and quantification of the in vitro activity of Aloe marlothii (A. Berger) subsp. marlothii and Elephantorrhiza elephantina (Burch.) skeels acetone extracts against Ehrlichia ruminantium.

An Ehrlichia ruminantium culture system was utilized for the anti-rickettsial evaluation of two ethnoveterinary plants, Elephantorrhiza elephantina and Aloe marlothii. Well-established E. ruminantium cultures were incubated with the plant leaf acetone extracts and compared to oxytetracycline and untreated controls. Effectivity was established by comparing the percentage parasitised cells and the calculation of both EC50 and extrapolated EC90 in microg/ml. The plant extracts were also screened for antibacterial activity using bioautography. Elephantorrhiza elephantina and A. marlothii demonstrated anti-ehrlichial activity with an EC50 of 111.4 and 64.5 microg/ml and EC90 of 228.9 and 129.9 microg/ml, respectively. The corresponding EC50 and EC90 for oxytetracycline was 0.29 and 0.08 microg/ml. Both plants appeared to produce their inhibitory activity by a similar mechanism, unrelated to that of the tetracyclines. Both the plant acetone extracts demonstrated antibacterial activity against Escherichia coli and Staphylococcus aureus (ATCC strains).

Effect of dimethyl sulfoxide in the treatment of sheep experimentally infected with Ehrlichia ruminantium.

OBJECTIVE: To evaluate the clinical response of sheep experimentally infected with Ehrlichia ruminantium to treatment with dimethyl sulfoxide (DMSO). ANIMALS: 32 Merino crossbred sheep. PROCEDURES: 16 sheep were infected with E ruminantium; 8 of these were treated twice daily with a 10% solution of DMSO (1 g/kg, i.v.) in polyionic fluid for 3 consecutive days. Treatment was initiated 2 days after the onset of clinical disease. Eight uninfected control sheep were similarly treated with DMSO. Placebo treatments (polyionic fluid administrations) were given to 8 infected and 8 uninfected sheep. Arterial and venous blood samples for blood gas and total plasma protein concentration measurements were collected daily (data from 5 days before until 6 days after onset of clinical disease were analyzed); physiologic variables and food consumption were also monitored. Gross pathologic findings and cytologic confirmation of the disease were recorded for the 16 infected sheep. RESULTS: Infected sheep treated with DMSO were able to maintain pulmonary gas exchange and had reduced pleural effusion and plasma protein loss, compared with infected untreated sheep that became hypoxic. Infected treated sheep developed an uncompensated metabolic acidosis. Uninfected treated sheep had reduced appetite, whereas uninfected untreated sheep maintained normal food intake. CONCLUSIONS AND CLINICAL RELEVANCE: Results of DMSO treatment in sheep with experimentally induced heartwater disease indicated that administration of this agent, in combination with specific antimicrobial treatment, may be of some benefit in treatment of naturally occurring disease.

Treatment of heartwater: potential adverse effects of furosemide administration on certain homeostatic parameters in normal sheep.

Diuretics, in particular furosemide, are generally recommended as a supportive treatment in the advanced stages of heartwater in ruminants. However, after what appeared to be possible adverse effects accompanying its use in field cases of heartwater, the effects of this drug on certain blood and urine parameters were investigated in normal sheep at the same dose rates. Diuresis with concomitant natriuresis was significant after furosemide administration, as was the expected plasma volume decrease. Other significant changes included metabolic alkalosis, hypokalaemia and reduced blood ionised calcium. The difference in duration of the diuretic effect and the duration of the changes in blood parameters from c. 3 h and c. 6 h respectively make it difficult to determine a time interval between successive treatments with furosemide. It appears that the probable cause of death of sheep with heartwater is a drastic reduction in blood volume and decreased cardiac output that leads to general circulatory failure. A therapeutic approach that involves further loss of plasma volume due to diuresis appears contradictory. The added effects of potentiating respiratory alkalosis and the terminal drop in blood ionised calcium seen in heartwater-affected animals indicate that the use of furosemide in supportive treatment of this disease is not warranted.

Heartwater in sheep and goats: a review.

Heartwater (cowdriosis) is an important, often fatal, tick-borne disease of domestic and wild ruminants in sub-Saharan Africa and some Indian Ocean and Caribbean islands. The causal agent, Cowdria ruminantium (Cowdry 1925), is a rickettsia closely related to members of the genus Ehrlichia, and is probably a part of a complex of genomic species. Imported breeds of sheep and goats (especially Angoras) are highly susceptible, but indigenous populations of endemic areas may be resistant to infection. Very young stock (less than 9 d old) possess a natural resistance that is unrelated to the immune status of the dams. Symptoms of heartwater vary, but usually begin with fever and may involve neurological signs and respiratory distress. Clinical diagnosis is based on symptoms, history of tick-exposure and post-mortem findings, and is confirmed by demonstration of characteristic rickettsial organisms in vascular endothelial cells. Laboratory diagnosis is retrospective and includes fluorescent antibody and enzyme-linked immunosorbent assays. Serological tests are compromised by non-specific reactions with certain Ehrlichia spp. DNA and oligonucleotide probes have been developed, but are thus far unavailable in many countries affected by heartwater. Treatment with tetracyclines is effective if begun in the early stages of infection. Control is based on a knowledge of the disease cycle in nature, and is achieved through judicious tick control, vaccination or both. A virulent, blood-based vaccine is available. Existence of a carrier state in recovered animals, including wild ruminants, complicates control efforts, and eradication is feasible only in circumscribed foci. Problem areas in fundamental and applied research on heartwater, as it affects sheep and goats, are discussed.

Chemotherapy and delivery systems: haemoparasites.

Chemotherapy of haemoparasitic diseases in domestic animals is dependent on a limited number of compounds, many of which are chemically closely related. In this review, a summary is given of each of the drugs currently available for treatment and prophylaxis of trypanosomosis and the tick-borne diseases theileriosis, babesiosis, anaplasmosis and cowdriosis. In contrast to the situation with the drugs used for tick-borne diseases, drug resistance appears to be becoming an increasing problem associated with the compounds used for trypanosomosis. The literature that has been reviewed, therefore, is that which relates to the methods used to identify and quantify drug resistance in trypanosome populations, reports of resistance to trypanocides, and cross-resistance between trypanosome populations, reports of resistance to trypanocides, and cross-resistance between trypanocides. The possible reason(s) for the apparent lack of development of resistance to the compounds used for treatment of tick-borne diseases is also discussed. Local toxicity at the site of injection is a problem that is particularly associated with many of the trypanocides when used on a long-term basis in individual animals. Various alternative preparations of the currently used trypanocides therefore have been evaluated in an attempt to reduce this toxicity, and are summarised. Finally, future developments in haemoparasitic chemotherapy are considered and, for trypanosomosis, highlight the importance of integrating chemotherapeutic and chemoprophylactic programmes with control of the vector when drug resistance becomes a significant constraint.

Methods currently used for the control of heartwater: their validity and proposals for future control strategies.

Major advances made in the last few years in the knowledge of heartwater have not yet resulted in significant improvement of its prevention and control. Acaricide resistance and the high cost of chemical vector control stimulate research on alternative methods of vector control, such as immunization against the tick and the use of pheromones. Recently developed in vitro methods for the culture of Cowdria ruminantium may make it possible to arrive at safe recombinant or molecular vaccines. Specific chemotherapy may be improved by new laboratory screening methods, and studies on the pathogenesis of heartwater could lead to better non-specific treatment as well. Studies on the heritability of natural resistance to heartwater should determine whether or not practical selection and transfer of resistance is feasible. More research is also needed on the immunology and diagnosis of the disease and the factors involved in the resistance of young animals.

[Prevention of cowdriosis and cases of sheep diseases in the Niayes region of Senegal]. / Prophylaxie de la cowdriose et observations sur la pathologie ovine dans la région des Niayes au Sénégal.

Sheep originating from an heartwater-free area were immunized by the infection and treatment method, using long-acting oxytetracycline. The animals were then exposed, together with control sheep to natural infection in an enzootic area. Tick counts were made to monitor the infestation and establish a correlation between infestation by certain tick species, in particular Amblyomma variegatum, and the occurrence of cowdriosis. In some cases of death associated with ehrlichiosis and anaplasmosis in the group immunized against cowdriosis, groups of Cowdria ruminantium were found in smears of the cerebral cortex, which leaves question marks as to the real nature of immunity to cowdriosis. Furthermore, these losses show the necessity of an integrated approach to the control of diseases of livestock in a given area. The presence of Rhipicephalus e. evertsi, formerly absent from this area, was significant in this respect.

Protection of goats against Caribbean and African heartwater isolates by the Ball 3 heartwater vaccine.

Two groups of castrated male adult goats (three goats/group) were infected intravenously with the Ball 3 vaccine strain of Cowdria ruminantium and treated with long-acting oxytetracycline at the onset of clinical disease 10 days later. Five weeks post-vaccination one group was challenged with a Caribbean isolate (Gardel) and the other group with a West African isolate (Mali) of C. ruminantium. Non-vaccinated controls infected with either the Gardel or Mali isolate died. All of the vaccinated animals challenged with the Gardel strain reacted and survived. The three vaccinated animals challenged with the Mali isolate also reacted and two died of heartwater.

Development of a prophylactic regime using long-acting tetracycline for the control of redwater and heartwater in susceptible cattle moved into an endemic area.

Thirty young crossbred steers were purchased from a tick-borne disease-free farm in South Africa, shown to be serologically negative to Babesia bigemina/bovis and Cowdria ruminantium, and moved to an experimental farm where babesiosis and heartwater were known to be endemic. The animals were allowed to graze as a herd on tick-infested areas of the farm for 90 days from the beginning of April but were separated into three groups of 10 for treatment. One group received a prophylactic regime of two injections of 20 mg/kg of a long-acting oxytetracycline on days 7 and 14 after introduction, one received a regime of three similar injections on days 7, 14 and 21 and the other group remained untreated prophylactically. The animals were observed early each morning in a crush, rectal temperatures being taken daily and blood smears when clinical signs of disease occurred. When clinical disease was confirmed in individual animals appropriate therapeutic treatment was implemented. In addition tick counts were made weekly. All untreated control animals required treatment for redwater between days 23 and 32 and eight of them then contracted heartwater of which five died despite therapy. Four animals from the group receiving two injections of oxytetracycline and one receiving three injections also contracted redwater but rather later. During the initial 35 day assessment period it was noticed that the majority of Boophilus ticks failed to engorge on the oxytetracycline-treated animals. In the second phase of the trial theileriosis (T. mutans-type) became evident in many animals and appeared to precipitate new or recrudescent clinical redwater and heartwater in several of them.(ABSTRACT TRUNCATED AT 250 WORDS)

The treatment of heartwater.

This paper reviews the available literature on the treatment of heartwater and draws comparisons with results obtained from recent work on experimentally-induced heartwater in sheep, calves and mice.

Identification of drugs effective against Cowdria ruminantium using a mouse screening system.

Heartwater is a tick-transmitted rickettsial infection of ruminants, caused by Cowdria ruminantium. The tetracyclines are the only compounds available for therapy of the disease. A screen, using mice infected with C. ruminantium, was developed and used to identify new compounds with potential for the control of heartwater. A series of di-4-methyl-thiosemicarbazones was shown to be highly active in the mouse model and their efficacy was confirmed in further trials in sheep infected with C. ruminantium. The mouse screen was shown to be simple to operate and reliably predictive of activity against heartwater. Ways in which the screen may be improved are suggested.

Observations on mouse-infective stocks of Cowdria ruminantium: attempts to demonstrate persistence of the organism in mice immune to the Kwanyanga stock.

Mice immunised against the Kwanyanga stock of Cowdria ruminantium by infection and treated with oxytetracycline proved immune to challenge on day 40 and also to a second challenge on day 125 after infection. Treatment with the experimental dithiosemicarbazone gloxazone on days 59 and 73 did not abolish immunity to challenge on day 125. No persistence of the organism in immune mice that had been challenged on day 40 could be demonstrated by subinoculating blood and liver homogenate on day 126. These results are different from findings reported elsewhere with the mouse-infective Kumm stock.

Heartwater in Angora goats. II. A pathological study of artificially infected, treated and untreated goats.

Pathological lesions in untreated Angora goats infected with the Ball3 strain of Cowdria ruminantium corresponded with those previously reported. A severe nephrosis was the most prominent pathological lesion in the animals treated after the 1st day of the febrile reaction. Renal ischaemia appears to be central to the pathogenesis of the kidney lesions.

Virulence of two strains of Cowdria ruminantium in mice and their use to predict drug activity against heartwater.

A study was made of the infectivity of two mouse-adapted strains of Cowdria ruminantium in mice. The Kwanyanga strain was most virulent in Balb/C mice which died nine days after infection with homogenate of liver from infected mice. CD-1 mice were least susceptible of six strains tested. The du Plessis strain of C. ruminantium was equally virulent in all six mouse strains. The du Plessis strain in CD-1 mice was used as the basis of a drug screen to detect activity against heartwater (C. ruminantium infection) and was highly predictive when active compounds were tested in sheep infected with the Ball 3 strain of C. ruminantium.

Cross immunity between strains of Cowdria ruminantium.

Cross immunity tests with strains of Cowdria ruminantium from South Africa, the Sudan, São Tomé and Nigeria failed to demonstrate antigenic differences. The antibiotic ampicillin showed no activity against heartwater.