Characteristics of Animal-assisted Interventions in the state of Lower Saxony, Germany, with a focus on hygiene in health care facilities.

Animal-assisted Interventions (AAIs) are becoming increasingly popular. To date, information on the extent of AAIs in Germany is limited. With a focus on infection control measures in health care facilities (HCFs), two studies were conducted in Lower Saxony to gain knowledge about the structure, characteristics and frequency of AAIs. An online survey among AAI providers identified dogs as the most important animal species in AAI, which mainly operated in educational facilities (53%) and/or on own property (46%). Twenty-nine percent offered their services in HCFs. The majority (55%) of the animal handlers was highly trained in AAI, but their awareness of hygiene and infection control measures to prevent zoonotic disease transmission was limited. Nineteen percent of animal handlers dewormed dogs only when faecal examinations were positive and 13% of dogs received ectoparasiticides only when infestations were present. Raw meat diets were frequent (82%). There was little awareness among animal handlers about the possibility of a zoonotic transmission from the client to the animal. Thus, handling of therapy dogs often reflected that of a "normal" pet ownership and did not always account for the special situation in HCFs. A telephone survey in 148 hospitals showed that 28% of the hospitals had experiences with animal-assisted therapies or animal visits, but 22% of these were lacking regulations on handling these animal contacts. While 28% of all hospitals had regulations for assistance dogs only 5% were aware of a new law that grants people accompanied by an assistance dog broad admission rights to public spaces, including HCFs. With an expected further increase in popularity of AAIs high quality standards which include infection control measures and animal welfare should be adopted by all AAI providers and recipients. This will ensure a safe implementation of this complementary medicine, where both sides - the human and the animal - can benefit.

10-year parasitological examination results (2003 to 2012) of faecal samples from horses, ruminants, pigs, dogs, cats, rabbits and hedgehogs.

The results of coproscopical examinations in domestic animals and hedgehogs carried out as routine diagnostics in the years 2003 to 2012 at the Institute for Parasitology, University of Veterinary Medicine Hannover, Germany, are presented. Of 3475 horse faecal samples, 30.1% contained stages of strongyles and 1.3% eggs of Strongyloides westeri and Parascaris equorum, respectively. The most frequently observed parasite stages in 1416 cattle faecal samples were Eimeria oocysts (21.3%) and strongyle eggs or larvae (15.9%). Dictyocaulus viviparus larvae and Fasciola hepatica eggs were identified in 0.9 and 1.3% of samples. Of 574 bovine faecal samples analysed by carbol-fuchsin staining, 39.9% were positive for Cryptosporidium oocysts. Stages of strongyles were found in 52.4% of sheep (n = 374) and 44.9% of goat faeces (n = 98) and Eimeria oocysts in 41.4 and 32.7% of their faeces, respectively. Of 1848 pig faecal samples, 3.0% contained stages of strongyles, 1.6% eggs of Ascaris suum and 3.3% coccidian (Eimeria or Cystoisospora spp.) oocysts. The most frequently detected helminth eggs in faecal samples of dogs (n = 2731) and cats (n = 903) were Toxocara spp. (2.8 and 3.9%, respectively). Cystoisospora oocysts were identified in 5.6% of dog and 2.4% of cat faeces. Furthermore, 0.7% of the cat samples were positive for small Toxoplasma gondii-like oocysts. The faecal samples of rabbits (n = 434) contained eggs of Passalurus ambiguus (3.0%), strongyles (1.8%) and Trichuris leporis (0.2%) as well as Eimeria oocysts (21.2%). The most abundant nematodes in the samples of hedgehogs (n = 205) were Capillaria spp. (39.5%) and Crenosoma striatum (26.8%); coccidian oocysts were found in 14.2% of the samples.

Chemical Compatibility and Safety of Imidacloprid/Flumethrin Collar (Seresto®) Concomitantly Used with Imidacloprid/Moxidectin (Advocate®, Advantage® Multi) and Emodepside/Praziquantel (Profender®) Spot-on Formulations.

Safety of concomitant use of veterinary products is of clinical interest. A series of studies was performed to evaluate the chemical compatibility and short term dermal and systemic safety of an imidacloprid/flumethrin collar (Seresto(®)/ Foresto(®), Bayer) used concomitantly with spot-on or tablet formulations.Chemical compatibility was evaluated in-vitro (study reference A) on collar pieces, followed by two small, non-controlled clinical studies (study reference B) in both, cats and dogs. The studies showed, that certain solvents affected the collar in-vitro, but not in their marketed formulations.Dermal and systemic safety of different spot-on or tablet formulations was first evaluated in a small, non-controlled clinical study (study reference C) in cats and dogs, via clinical observations only, followed by controlled clinical safety studies of concomitant use with imidacloprid/ moxidectin (Advocate(®)/ Advantage(®) Multi, Bayer) in dogs and cats (study reference D) and emodepside/ praziquantel (Profender(®), Bayer) in cats (study reference E), assessing safety aspects by clinical observations and statistical analyses of hematology and clinical chemistry parameters compared to baseline values and between treated and control groups.Dermal safety findings over all clinical studies (study references B to E) matched those already described for the respective products and included transient cosmetic changes (oily hair and crystal formation) at the site of spot-on application and broken hair, transient alopecia and skin alterations at the site of collar application. There were no indications of these findings aggravating under the conditions of concurrent use. There were no systemic safety findings of clinical significance in any of the clinical safety studies (study reference C to E). Assessment of blood parameters revealed some deviations from baseline levels and from the reference range in dogs as well as in cats, but no clinical relevance could be deduced. Hematology and clinical chemistry results confirmed the safety of the concomitant treatment. It is concluded that Seresto(®) is chemically compatible with solvents used in major spot-on formulations on the market and is dermally and systemically safe for adult dogs and cats when used concomitantly with Advocate(®) and Profender(®) spot-on formulations.

Efficacy of Emodepside/Praziquantel Spot-on (Profender®) against adult Aelurostrongylus abstrusus Nematodes in Experimentally Infected Cats.

The adulticidal efficacy of a topical combination of emodepside 2.1 % (w/v) plus praziquantel 8.6 % (w/v) (Profender® spot-on for cats, Bayer) against adult Aelurostrongylus abstrusus nematodes was evaluated in two randomised, placebo-controlled laboratory efficacy studies. Each study involved 16 cats experimentally inoculated with L3 (800 and 600 each in studies no. 1 and 2, respectively) and randomised into two study groups of 8 cats each after onset of patency. While cats in the treatment group in study no. 1 received a single spot-on application at the minimum therapeutic dose (3 mg/kg emodepside and 12 mg/kg praziquantel), cats in study no. 2 were treated twice with an interval of 14 days. The faecal output of first stage larvae was monitored throughout the study. Necropsy was conducted 4 or 5 weeks after the (first) treatment and the worm counts were used for efficacy calculations. The control groups showed a geometric mean of the total worm count (live and dead worms) of 28.8 (study no. 1) and 17.6 (study no. 2), respectively. All control animals were infected. While the single treatment in study no. 1 resulted in a reduction of the total worm burden by 73.0 % (p = 0.0070), the treatment protocol in study no. 2 was 99.2 % effective (p = 0.0035). Based on live worm counts, the efficacy in study no. 2 was 100 % (p = 0.0030). It is concluded that two applications of Profender® spot-on given two weeks apart represent a safe and highly efficacious treatment regime against feline aelurostrongylosis.

Prevention of Lactogenic Toxocara cati Infections in Kittens by Application of an Emodepside/Praziquantel Spot-on (Profender®) to the Pregnant Queen.

This study aimed to evaluate the efficacy of an emodepside 2.1 % (w/v)/praziquantel 8.6 % (w/v) topical solution (Profender® spot-on for cats) in the prevention of lactogenic Toxocara cati infections. A controlled test was performed with two groups of 8 cats with confirmed pregnancy. All cats were infected with daily doses of 2000 T. cati eggs for 10 consecutive days starting 50 days post conception to produce an acute infection. Treatment was performed 60 days post conception. Queens in the treatment group received the emodepside/praziquantel solution at the minimum therapeutic dose (3 mg/kg emodepside and 12 mg/kg praziquantel), while the control group was treated with a placebo spot-on. Efficacy was evaluated 56 days post partum by necropsy of one randomly selected kitten of each litter and comparison of the worm burdens between the study groups. Additionally the necropsy results were supported by quantification of worms expelled with the faeces after deworming of the remaining kittens and all queens. The treatment in late pregnancy resulted in an efficacy of 98.7 % (p < 0.0001). All necropsied control kittens were infected (geometric mean 30.6). Seven of 8 kittens from treated mothers were free of T. cati (geometric mean 0.4). Worm counts after deworming reflected the results obtained at necropsy. No side effects of the treatment were observed. It is concluded that treatment with an emodepside/praziquantel spot-on solution during late pregnancy effectively prevents lactogenic transmission of T. cati to the offspring. The study design facilitated the generation of reliable data, while at the same time a minimum number of animals was sacrificed.

Enoxaparin: pharmacokinetics and treatment schedule for cats.

Detailed pharmacokinetic data are not available for subcutaneously (SC) administered enoxaparin in cats and this causes difficulties in establishing treatment protocols. The aims of this study were (1) to establish pharmacokinetic data of SC administered enoxaparin and (2) to establish a treatment schedule. Six healthy cats received a single SC injection of 1 mg enoxaparin/kg and blood samples were collected before and 1, 2, 3, 4, 6, 8, 10 and 12 h after the injection. Six further healthy cats received 0.75 mg/kg every 6 h for four consecutive days and blood samples were collected before and 2 h after the first and second injection on day 1, and the first injection on days 2 and 4. Anti-factor Xa (FXa) activity, coagulation tests and thromboelastometry assays were performed. Enoxaparin injection was well tolerated. Following the single SC injection Cmax was 0.83 ± 0.08 anti-Xa IU/mL and in 5/6 cats was detected after 2 h (Tmax = 110 ± 25 min). The total clearance was 23.4 ± 4.8 mL/h/kg and the terminal half-life was 2.27 ± 0.4 h. All cats receiving repeated injections reached the defined target peak range of 0.5-1.0 IU/mL by 2 h after the second injection (0.54 [0.50-0.61]; median, [minimum - maximum]) and there was no considerable accumulation subsequently. With the exception of thromboelastometry (especially non-activated), ratio values of coagulation times increased significantly although only slightly (e.g., the maximal value of median activated partial thromboplastin time ratio was 1.27). Significant, although only moderately close relationships with Spearman rank correlation coefficients between 0.424 and 0.558 were calculated between anti-FXa activities and ratios of different coagulation times. A dosage schedule of 0.75 mg/kg four times a day seems suitable for therapeutic use of enoxaparin in cats as it leads to reproducible peak anti-FXa activities within the target range for the treatment of thrombosis in humans. The low inter-individual variation may indicate that monitoring based on anti-FXa activities is not necessary.

Assessment of the combination of spinosad and milbemycin oxime in preventing the development of canine Angiostrongylus vasorum infections.

Angiostrongylus vasorum is an increasingly reported parasite in Europe that develops in dogs after ingestion of infective third stage larvae (L3) that reside in gastropod molluscs which are needed to complete the parasite's life-cycle. Infection can produce a diversity of clinical signs, determined by involvement of the respiratory, neurological, and/or coagulation system, with a likely fatal outcome in the absence of treatment. Few drugs have been shown to reliably prevent infection, and data on treatment of infections is limited. A controlled, randomized, partially blinded laboratory study was therefore executed to evaluate the efficacy and safety of a combination tablet of spinosad/milbemycin oxime in dogs inoculated with approximately 250 A. vasorum L3. Sixteen healthy nematode free adult dogs were randomly allocated to two study groups of 8 dogs each. Thirty days post inoculation (dpi) all dogs in the fed state were treated: dogs in group B were treated with spinosad and milbemycin oxime at the dose rates of 45-60 mg/kg and 0.75-1.0mg/kg bodyweight, respectively, approximately the lower half portion of the expected full unit dose range; dogs in group A were treated with placebo tablets. All dogs were euthanized and necropsied 56-58 dpi. The heart and lungs were examined to determine the presence of A. vasorum. All placebo group dogs were infected at necropsy with counts ranging from 22 to 98 adult worms and a geometric mean worm count of 55.2. In contrast, the geometric mean worm count in the spinosad/milbemycin oxime group was 0.7 with worm numbers ranging from 0 to 8. The results of this study demonstrate that a single treatment with the tablet combination of spinosad and milbemycin oxime administered 30 dpi provided 98.8% preventive efficacy against development of adult A. vasorum infections. Monthly treatments with spinosad and milbemycin oxime have the potential to prevent the establishment of infections with A. vasorum in dogs.

Efficacy of emodepside plus toltrazuril oral suspension for dogs (Procox®, Bayer) against Trichuris vulpis in naturally infected dogs.

The efficacy of emodepside plus toltrazuril oral suspension for dogs (Procox®, Bayer) against Trichuris vulpis was evaluated in a controlled, blinded and randomised laboratory study. Twenty naturally infected dogs were included. Dogs in the treatment group received the minimum therapeutic dose of 0.45 mg emodepside and 9 mg toltrazuril per kg body weight, while dogs in the control group were left untreated. Efficacy was calculated based on worm counts after necropsy on Day 7 post treatment. Additionally, all faeces were collected and examined for expelled worms. The treatment was 100 % effective. A total of 233 adult worms (geometric mean 17.0) and 3 immature adult worms were found in the control group at necropsy. Adequacy of infection was demonstrated. The treated group excreted a total of 186 adult worms within 2 days after treatment. Additionally, all dogs were co-infected with Uncinaria stenocephala. Efficacy against this parasite was 99.8 %. No side effects of the treatment were observed. This study demonstrates that in addition to the formerly proven efficacy against Toxocara canis, Ancylostoma caninum and Uncinaria stenocephala, emodepside plus toltrazuril suspension is also effective against T. vulpis and thus represents a convenient treatment option for dogs co-infected with whipworms and coccidia.

Discrimination of gastrointestinal nematode eggs from crude fecal egg preparations by inhibitor-resistant conventional and real-time PCR.

Diagnosis of gastrointestinal nematodes relies predominantly on coproscopic methods such as flotation, Kato-Katz, McMaster or FLOTAC. Although FLOTAC allows accurate quantification, many nematode eggs can only be differentiated to genus or family level. Several molecular diagnostic tools discriminating closely related species suffer from high costs for DNA isolation from feces and limited sensitivity since most kits use only small amounts of feces (<1 g). A direct PCR from crude egg preparations was designed for full compatibility with FLOTAC to accurately quantify eggs per gram feces (epg) and determine species composition. Eggs were recovered from the flotation solution and concentrated by sieving. Lysis was achieved by repeated boiling and freezing cycles - only Trichuris eggs required additional mechanic disruption. Egg lysates were directly used as template for PCR with Phusion DNA polymerase which is particularly resistant to PCR inhibitors. Qualitative results were obtained with feces of goats, cattle, horses, swine, cats, dogs and mice. The finally established protocol was also compatible with quantitative real-time PCR in the presence of EvaGreen and no PCR inhibition was detectable when extracts were diluted at least fourfold. Sensitivity was comparable to DNA isolation protocols and spiked samples with five epg were reliably detected. For Toxocara cati a detection limit below one epg was demonstrated. It was possible to distinguish T. cati and Toxocara canis using high resolution melt (HRM) analysis, a rapid tool for species identification. In human samples, restriction fragment length polymorphism (RFLP) and HRM analysis were used to discriminate Necator americanus and Ancylostoma duodenale. The method is able to significantly improve molecular diagnosis of gastrointestinal nematodes by increasing speed and sensitivity while decreasing overall costs. For identification of species or resistance alleles, analysis of PCR products with many different post PCR methods can be used such as RFLP, reverse-line-blot, Sanger sequencing and HRM.

An immunoblot for detection of Taenia saginata cysticercosis.

Control measures to prevent human infections with the food-borne zoonotic helminth Taenia saginata are currently based on meat inspection, which shows rather low diagnostic sensitivity. To develop an immunoblot for detection of T. saginata-infected cattle, crude proteins of T. saginata cysts were extracted and separated with SDS-PAGE. The cyst antigens showed ten protein bands ranging from 260 to 14 kDa. T. saginata cyst proteins 260, 150, 130, 67, 60, 55, 50, and 23 kDa were immunoreactive with known positive sera of T. saginata-infected cattle but cross-reacted with sera from Echinocccus granulosus-infected ruminants. By contrast, 14- and 18-kDa cyst proteins reacted specifically with T. saginata-positive sera and thus might be potential candidates for the development of a T. saginata-specific immunoassay. Proteins of E. granulosus cysts and Taenia hydatigena cysts were also extracted and separated with SDS-PAGE. E. granulosus cysts revealed 11 protein bands ranging from 260 to 23 kDa. E. granulosus protein 60 kDa was immunoreactive with E. granulosus-positive sera only. The cyst of T. hydatigena showed 11 protein bands ranging from 290 to 14 kDa. The protein band 35 kDa showed cross-reaction with positive sera from both T. saginata- and E. granulosus-infected animals. A protein of 67 kDa was present in all three tested cestode species and was the major antigenic protein detected by sera of T. saginata- and E. granulosus-infected animals. Therefore, this protein represents a potential vaccine candidate against both cysticercosis and cystic echinococcosis in cattle.

Treatment of third-stage larvae of Toxocara cati with milbemycin oxime plus praziquantel tablets and emodepside plus praziquantel spot-on formulation in experimentally infected cats.

Toxocara cati is the most prevalent gastrointestinal helminth in cats worldwide, with cats of all ages at risk of infection. An anthelminthic treatment that not only affects the gut-dwelling stages of this parasite but is also effective against developmental stages in the tissue has the advantage that the pathology caused by migrating larvae is minimized and the need for repeated treatments is reduced. This study was conducted to evaluate the efficacy of milbemycin oxime/praziquantel tablets (Milbemax®, Novartis) against third-stage larvae of T. cati in comparison to a spot-on formulation of emodepside and praziquantel (Profender®, Bayer). Twenty-four kittens were experimentally infected with T. cati and randomly allocated to three study groups. Treatments were performed at the minimum therapeutic dosage 5 days after the experimental infection. The development of patent infections was monitored and all cats were dewormed 50 days post-infection. Efficacies were calculated based on counts of excreted worms in the treated groups compared to a negative control group. Seven of the eight cats in the negative control group developed a patent T. cati infection and all cats were excreting worms at the end of the study (geometric mean worm count 18.1). No efficacy could be observed for the milbemycin oxime-treated animals. All cats developed a patent infection and excreted worms (geometric mean worm count 27.7). The treatment with Profender® was 98.5 % effective against L3 of T. cati. One cat developed a patent infection and was excreting worms at the end of the study (geometric mean worm count 0.3). No adverse reactions were noted in either treatment group.

Pharmacokinetics of the low molecular weight heparin dalteparin in cats.

Low molecular weight heparin (LMWH) is used as an anticoagulant in cats although only limited pharmacokinetic data are available in this species. The aim of the present study was to establish the pharmacokinetics of dalteparin in cats based on anti-FXa heparin activities. Groups of clinically healthy cats (six animals per treatment) received individual LMWH injections at three different doses intravenously (IV) (25, 50, 100 anti-factor Xa international units [IU anti-FXa]/kg) or subcutaneously (SC) (50, 100, 200 IU anti-FXa/kg). Blood samples were collected before and at various times after injection. Anti-FXa activity was measured with a chromogenic substrate test. Following IV injection, maximum plasma heparin activities (C(max)) were 0.67 ± 0.14, 1.44 ± 0.22 and 2.87 ± 0.38 IU anti-FXa/mL, respectively. The calculated mean half-life (t(½)) was between 39 and 57 min and was not significantly dose-dependent (P=0.139). The volume of distribution (35-39 mL/kg) was almost equivalent to the plasma volume. After SC injection, C(max) values of 0.41 ± 0.10, 0.86 ± 0.17 or 1.91 ± 0.16 IU anti-FXa/mL, respectively, were calculated at 91-110 min post-injection. The t(½) values were between 106 and 122 min and were not significantly influenced by dose (P=0.784). The bioavailability after SC injection was approximately 100%. The high bioavailability of the SC administered LMWH dalteparin in cats was consistent with other species and indicated predictable blood levels. However, the comparatively short t(½) may indicate the necessity of multiple daily injections, which should be verified in clinical trials.

Efficacy of emodepside plus toltrazuril (Procox(®) oral suspension for dogs) against Toxocara canis, Uncinaria stenocephala and Ancylostoma caninum in dogs.

The efficacy of emodepside plus toltrazuril (Procox® oral suspension for dogs) against different species of gastrointestinal nematodes (Toxocara canis, Ancylostoma caninum, Uncinaria stenocephala) was evaluated in nine randomised,blinded and placebo-controlled laboratory studies in naturally or experimentally infected dogs. The product was used at the proposed minimum dose of 0.45 mg emodepside and 9 mg toltrazuril per kg body weight. Efficacy was calculated based on worm counts after necropsy. Worm burdens in the control dogs ranged between 0 and 409 worms of the respective stage for T. canis and between 4 and 655 worms for hookworms. The studies demonstrated 100 % efficacy of emodepside/toltrazuril suspension against mature adult, ≥ 94.7 %efficacy against immature adult and 99.3 % efficacy against the L4 larval stage of T. canis. The efficacy against mature adult A. caninum was ≥ 99.5 % and the efficacy against mature adult U. stenocephala was 100 %. All differences between treatment and control groups were statistically significant and no gender effect was found. It can be concluded that the emodepside/toltrazuril suspension represents a safe and highly effective product in dogs with nematode (T. canis, hookworms) infection.

Dynamics of distribution and efficacy of different spot-on permethrin formulations in dogs artificially infested with Dermacentor reticulatus.

BACKGROUND: Varying reports concerning the duration and reliability of different permethrin preparations' efficacy can be found in the literature. The aim of this study was to investigate the dynamics of the distribution and efficacy of four different spot-on formulations with permethrin as the active ingredient formulated with different solvents. To examine the influence of these solvents on the speed of distribution and the acaricidal activity of permethrin in the coat, an in vivo study under laboratory conditions was performed. Six dogs per test period were treated with the recommended dose and 1, 14 and 28 days after treatment dogs were infested with Dermacentor reticulatus ticks: a) on the back, near the application site, and b) on the hind leg, the greatest possible distance from the application site. Efficacies were determined 6 hours after tick infestation to examine the repellent effect and the speed of kill of the products which plays an important role in the context of tick transmitted diseases. RESULTS: After six hours of exposure, a significant acaricidal efficacy (p < 0.001) could be observed in all treated groups over the whole duration of the study, regardless of which product was used. While the arithmetic mean of attached ticks was < 3 on Day 1, numbers increased over the course of the study to a mean of > 9 on Day 28. However, most of these ticks were dead even 28 days after treatment, as the mean of live attached ticks was still < 2. Significant differences could neither be found between the different permethrin spot-on formulations, nor between the two parts of the body (p > 0.05). CONCLUSIONS: All products were able to kill ticks within six hours following infestation from Day 1 to Day 28 after treatment. Additionally, no significant difference between the tick numbers on the back and the hind leg could be found at any time, which implies a homogenous distribution of permethrin over the body. The efficacy of all four products was comparable during the whole study period, showing that the different solvents do not significantly affect the dynamics of distribution.

Efficacy of emodepside plus praziquantel tablets (Profender tablets for dogs) against mature and immature infections with Toxocara canis and Toxascaris leonina in dogs.

The efficacy of emodepside plus praziquantel tablets (Profender tablets for dogs) against mature adult, immature adult and larval stages of Toxocara canis and Toxascaris leonina was evaluated in ten randomised, blinded and placebo-controlled dose confirmation studies in naturally or experimentally infected dogs. The tablets were used at the proposed minimum dose of 1 mg emodepside and 5 mg praziquantel per kg body weight. Efficacy was calculated based on worm counts after necropsy. Five studies demonstrated >99% efficacy against mature adult, >92% efficacy against immature adult, >98% efficacy against L4 and >94% efficacy against L3 larval stages of T. canis. Another five studies demonstrated >99% efficacy against mature and immature adult and >95% efficacy against L4 larval stages of T. leonina. No side effects of the treatment were observed. Emodepside plus praziquantel tablets thus provide a comprehensive new treatment option for ascarid infections in the dog.

Treatment and prevention of vertical transmission of Toxocara cati in cats with an emodepside/praziquantel spot-on formulation.

This study aimed to determine the efficacy of emodepside 2.14%/praziquantel 8.58% topical solution (Profender, Bayer) in the prevention and treatment of lactogenic Toxocara cati infections. Eight pregnant cats were orally infected with T. cati eggs during late pregnancy. Four queens were treated on day 60 post conception and four queens were left untreated. The kittens of two untreated queens were treated 28 days after birth. The two other negative control litters were left untreated. The efficacy of emodepside was determined by faecal egg counts. While faecal samples of queens and litters in the control group became positive for T. cati, egg shedding was completely prevented in all four treated queens, in their litters and in the kittens from the two litters which were treated four weeks after birth. The untreated mothers of the latter stayed also coproscopically negative, which might be explained by an oral uptake of emodepside through grooming. The treatment was well tolerated by pregnant queens as well as by four-weeks-old kittens.To our knowledge, this is the first publication that focuses on the prevention of lactogenic transmission of T. cati.

Evaluation of reference genes for quantitative real-time PCR to investigate protein disulfide isomerase transcription pattern in the bovine lungworm Dictyocaulus viviparus.

Quantitative real-time PCR (qPCR) is the most sensitive technique for transcript quantification provided that gene transcription patterns are normalized to an evaluated reference gene. For Dictyocaulus viviparus, the housekeeping genes beta-tubulin, beta-actin, elongation factor 1alpha (ef-1alpha), glyceraldehyde-3-phosphatase dehydrogenase (gapdh), and 60S ribosomal protein L37a (60S rpL37a) were characterized and evaluated. Evaluation using the geNorm software revealed ef-1alpha and beta-tubulin as the most suitable reference genes, whereas the coefficient of variation approach resulted in ef-1alpha and 60S rpL37a as transcripts with the least variation among 12 developmental lungworm stages. The critical influence of reference genes on qPCR data analysis, with the possible consequence of erroneous, misleading results due to inappropriate reference genes used for data normalization, is shown for protein disulfide isomerase 2 (pdi-2) transcription patterns. Proper normalization of pdi-2 transcription using ef-1alpha and beta-tubulin as reference genes resulted in a more than 7-fold enriched pdi-2 transcription in L1 compared to that in eggs, and a dramatic decrease in L3. Following an increase in the L5 stage there is again a decrease of pdi-2 transcription in adult lungworms. These fluctuations in the transcription levels reflect the requirement of cuticule collagen during bovine lungworm development.