Vet. Sci., Volume 5, Issue 3 (September 2018) – 21 articles

Brucella infections in wildlife have gained a lot of interest from the scientific community and different stakeholders. These interests are often different and sometimes conflicting. As a result, different management perspectives and aims have been implemented (One Health, public health, veterinary public health, maintenance of a brucellosis free status in livestock, sustainable wildlife harvesting by hunters, wildlife and environmental health). When addressing Brucella infection in wildlife, the most important features of Brucella infection should be considered and the following questions need to be answered: (1) Is Brucella infection a result of a spillover from livestock or is it a sustainable infection in one or more wildlife host species? (2) Did the epidemiological situation of Brucella infection in wildlife change over time and, if so, what are the main drivers of change and does it impact the wildlife population dynamics? (3) Does Brucella infection in wildlife represent a reservoir of Brucella strains for livestock? (4) Is Brucella infection in wildlife of zoonotic concern? These questions point to the fundamental biological question of how animal (domestic and wildlife)/Brucella spp. partnerships are established. Will we be able to decipher an evolutionary trajectory through a livestock-to-wildlife “host jump”? Whole genome sequencing and new “omics” techniques will help in deciphering the molecular basis of Brucella host preference and open new avenues in brucellosis management aimed at preventing opportunities for Brucella host jumps. Full article

Opuntia ficus-indica (L.) Mill. is a xerophylous plant that originated in tropical and subtropical America. This plant is popularly known in Brazil as “palma forrageira” (cactus pear) and plays a fundamental role in animal nutrition, mainly in the Northeastern semi-arid region of the country. The plant has several uses since it presents bioactive compounds that confer biological and pharmacological properties. In this context, the cactus pear can also be considered a potential product to combat parasite infections. The objective of this study was to chemically characterize the O. ficus-indica hydroalcoholic extract (OFIEOH) and to determine its efficacy against gastrointestinal parasites using in vitro tests. Initially, the hydroalcoholic extract from cladode peels of O. ficus-indica was produced by maceration for 21 days. For the chemical characterization, colorimetric dosages were performed for carbohydrates, proteins, phenols and condensed tannins. Liquid chromatography coupled to mass spectrometry/electron spray ionization (LC-MS/ESI) was used to characterize the polyphenolic profile of the OFIEOH extract. Fifteen compounds were identified in the OFIEOH extract, such as methyl, glycosylated and aglycone quercetin derivatives and aglycone and glycosylated kaempferol derivatives. Tri-glycosylated methyl quercetin derivatives were the main compounds identified. In vitro egg hatch (EHT) and larval migration tests (LMT) were used in a range of concentrations of OFIEOH from 12.5 to 100 mg/mL for EHT and 12.5 to 200 mg/mL for LMT. In addition, the LMT was used to test ivermectin (IVM) (from 11.4 to 57.1 µM), associated with the inhibitory concentration of 50% (IC₅₀) for OFIEOH. The combination of OFIEOH (12.5 to 200 mg/mL) plus the IC₅₀ of IVM was also tested. The efficacy of OFIEOH alone varied from 19.33 to 90.0% using the EHT. The LMT revealed an efficacy of 5.78 to 77.26% for the extract. Both tests showed a concentration-dependence inhibitory effect. We found a drug-extract antagonistic neutralizing effect when doses of IVM were added to OFIEOH (maximum efficacy of 73.78%), while a positive additive effect was observed when OFIEOH was added to the IC₅₀ of IVM (IC₅₀ of 82.79 for OFIEOH alone against an IC₅₀ of 55.08 of OFIEOH + IVM). The data from this work indicate that OFIEOH alone may be considered as a suitable ecofriendly product to control gastrointestinal parasites of sheep, offering a more holistic approach to improve animal farming and welfare. The drug-extract interaction is also a promising therapeutic alternative, reducing the final dose to the host, with an optimum combination effect. Full article

Levels of Japanese cedar pollen (Cryptomeria japonica) have increased in Japan and cedar pollinosis caused by Japanese cedar pollen has been reported in dogs. Serum levels of immunoglobulin E (IgE) against Cry j 1 and Cry j 2 in dogs raised in institutes and treated at veterinary hospitals in Japan were thus investigated. A total of 71 sera obtained from two institutes and 87 sera obtained from veterinary hospitals in the Hyogo and Kanagawa Prefectures were analyzed in this study. Serum levels of IgE were measured using the enzyme-linked immunosorbent assay with commercial purified Cry j 1 and Cry j 2. IgE against Cry j 1 and Cry j 2 in sera obtained from the two institutes were detected, despite the dogs being bred in enclosed areas. Moreover, significant differences were noted in the serum levels of IgE against Cry j 1 and Cry j 2 between the two institutes. The number of samples showing Cry j 1 or Cry j 2 levels above the cut-off values was greater in the Kanagawa Prefecture than in the Hyogo Prefecture. In total, 14 dogs showed Cry j 1 and Cry j 2 levels greater than the cut-off values in the Hyogo Prefecture, and only three such dogs were seen in the Kanagawa Prefecture. A significant correlation between serum levels against both allergens was observed (r² = 0.6931, p < 0.0001). Full article

By using the egg hatch test (EHT), the larval development test (LDT) and the larval mortality/paralysis test (LMT), the in vitro anthelmintic activity on sheep gastrointestinal strongyles (GIS) of four plant-derived pure compounds, mangiferin (at 0.25%, 0.125% and 0.0625%), rutin (at 1%, 0.75%, 0.5%), quercetin (at 1%), and β-sitosterol (at 1%, 0.75%, 0.5%), was investigated. For comparison, untreated and treated (0.1% thiabendazole, 0.1% TBZ) controls were used. Six repetitions were made throughout the experiment. Data were statistically elaborated using the χ² test. The concentration able to inhibit the development of the 50% of L1s to L3s and causing the mortality of the 50% of L3s (EC50) was also calculated. L3s recovered from untreated Petri dishes were identified at the genus level. In EHT, all tested compounds at all concentrations significantly (p < 0.01) inhibited the hatch of the eggs when compared to the untreated controls, but none of them was as effective as 0.1% TBZ. In LDT, rutin (at 1%, 0.75% and 0.5%), mangiferin (at 0.25% and 0.125%), β-sitosterol (at 1%) and 0.1% TBZ completely prevented the larval development from L1 to L3 in respect to the untreated controls (p < 0.01). In LMT, all tested compounds significantly (p < 0.01) increased the death of L3s compared to the untreated controls, except for β-sitosterol at 0.5%. However, only rutin at all concentrations and 0.25% and 0.125% mangiferin were as effective as 0.1% TBZ. Haemonchus, Trichostrongylus, Chabertia and Teladorsagia/Ostertagia GIS genera, were identified. Full article

Estimates of dog population sizes are a prerequisite for delivering effective canine rabies control. However, dog population sizes are generally unknown in most rabies-endemic areas. Several approaches have been used to estimate dog populations but without rigorous evaluation. We compare post-vaccination transects, household surveys, and school-based surveys to determine which most precisely estimates dog population sizes. These methods were implemented across 28 districts in southeast Tanzania, in conjunction with mass dog vaccinations, covering a range of settings, livelihoods, and religious backgrounds. Transects were the most precise method, revealing highly variable patterns of dog ownership, with human/dog ratios ranging from 12.4:1 to 181.3:1 across districts. Both household and school-based surveys generated imprecise and, sometimes, inaccurate estimates, due to small sample sizes in relation to the heterogeneity in patterns of dog ownership. Transect data were subsequently used to develop a predictive model for estimating dog populations in districts lacking transect data. We predicted a dog population of 2,316,000 (95% CI 1,573,000–3,122,000) in Tanzania and an average human/dog ratio of 20.7:1. Our modelling approach has the potential to be applied to predicting dog population sizes in other areas where mass dog vaccinations are planned, given census and livelihood data. Furthermore, we recommend post-vaccination transects as a rapid and effective method to refine dog population estimates across large geographic areas and to guide dog vaccination programmes in settings with mostly free roaming dog populations. Full article

The genomes of influenza A viruses (IAVs) comprise eight negative-sense single-stranded RNA segments. In addition to the protein-coding region, each segment possesses 5′ and 3′ non-coding regions (NCR) that are important for transcription, replication and packaging. The NCRs contain both conserved and segment-specific sequences, and the impacts of variability in the NCRs are not completely understood. Full NCRs have been determined from some viruses, but a detailed analysis of potential variability in these regions among viruses from different host groups and locations has not been performed. To evaluate the degree of conservation in NCRs among different viruses, we sequenced the NCRs of IAVs isolated from different wild bird host groups (ducks, gulls and seabirds). We then extended our study to include NCRs available from the National Center for Biotechnology Information (NCBI) Influenza Virus Database, which allowed us to analyze a wider variety of host species and more HA and NA subtypes. We found that the amount of variability within the NCRs varies among segments, with the greatest variation found in the HA and NA and the least in the M and NS segments. Overall, variability in NCR sequences was correlated with the coding region phylogeny, suggesting vertical coevolution of the (coding sequence) CDS and NCR regions. Full article

Prolapsed nictitans gland (PNG) is an important ocular condition of dogs. Various surgical interventions have been described, but effective technique is currently considered to be a matter of personal clinician preference. The aim of this rapid review was to evaluate existing peer-reviewed evidence of effectiveness for surgical techniques and their subsequent effects on quantitative and clinical lacrimal outcomes for PNG. We performed a structured bibliographic search of CAB Abstracts, PubMed, and Medline using terms relevant to dogs, nictitans gland, and surgery on 13 September 2017. Included studies were assessed for study design, reporting characteristics, surgical techniques, and surgical and lacrimal outcomes. Fifteen of three hundred fifteen identified studies were eligible for inclusion. Seven different replacement techniques were identified, along with gland excision. All studies were observational or descriptive, with the exception of a single crossover trial. Outcomes reporting was heterogeneous and provided limited detail on lacrimal outcomes or on breed propensity for recurrence. Insufficient data precluded comparison of techniques for either surgical failure rates or lacrimal outcomes, although proportional meta-analysis yielded an overall failure rate of 3% (95% CI 1–7%) for the Morgan’s pocket procedure. Improved reporting of veterinary surgical studies will improve evidence appraisal and synthesis, as well as reduce potential sources of bias. Full article

The cattle tick Rhipicephalus microplus is one of the most important ectoparasites causing significant economic losses for the cattle industry. The major tool of control is reducing the number of ticks, applying acaricides in cattle. However, overuse has led to selection of resistant populations of R. microplus to most of these products, some even to more than one active principle. Thus, exploration for new molecules with acaricidal activity in R. microplus has become necessary. Triosephosphate isomerase (TIM) is an essential enzyme in R. microplus metabolism and could be an interesting target for the development of new methods for tick control. In this work, we screened 227 compounds, from our in-house chemo-library, against TIM from R. microplus. Four compounds (50, 98, 14, and 161) selectively inhibited this enzyme with IC₅₀ values between 25 and 50 μM. They were also able to diminish cellular viability of BME26 embryonic cells by more than 50% at 50 μM. A molecular docking study showed that the compounds bind in different regions of the protein; compound 14 interacts with the dimer interface. Furthermore, compound 14 affected the survival of partially engorged females, fed artificially, using the capillary technique. This molecule is simple, easy to produce, and important biological data—including toxicological information—are available for it. Our results imply a promising role for compound 14 as a prototype for development of a new acaricidal involving selective TIM inhibition. Full article

A two-year-old, intact female Scottish Terrier presented with one-and-a-half-year history of erosive and ulcerative lesions affecting the nasal planum. Clinical appearance, history, histopathology, and response to therapy were suggestive of a rare vasculopathy of the nasal planum that has been previously described in Scottish Terrier dogs. In previously published reports, medical treatments of the disease had failed, leading to euthanasia of five dogs, while a short-term follow-up was available for one case that was controlled with prednisolone and ciclosporin. The dog reported herein was successfully treated with medical therapy consisting initially of a combination of ciclosporin and prednisolone and endonasal stents applied over the first six months. Stents were inserted in order to prevent abnormal scarring and nostril stenosis. More than one and a half years after diagnosis, the dog is still being administered ciclosporin once daily, breathes normally, and has an optimal quality of life. Full article

Feline oral squamous cell carcinoma (OSCC) is a highly invasive form of cancer in cats. In human OSCC, cluster of differentiation 147 (CD147) contributes to inflammation and tumor invasiveness. CD147 is a potential therapeutic target, but the expression of CD147 in feline OSCC has not been examined. Immunohistochemistry was used to determine if cyclooxygenase 2 (COX-2) and CD147 expression in feline OSCC biopsies was coordinated. Tumor cells were more likely to express COX-2 (22/43 cases or 51%) compared to stroma (8/43 or 19%) and adjacent oral epithelium (9/31 cases or 29%) (p < 0.05). CD147 was also more likely to occur in tumor cells compared to stroma and adjacent mucosa, with 21/43 (49%) of cases having >50% tumor cells with mild or moderate CD147 expression, compared to 9/28 (32%) in adjacent epithelium and only 5/43 (12%) in adjacent stroma (p < 0.05). In feline OSCC cell lines (SCCF1, SCCF2, and SCCF3), CD147 gene expression was more consistently expressed compared to COX-2, which was 60-fold higher in SCCF2 cells compared to SCCF1 cells (p < 0.05). CD147 expression did not correlate with COX-2 expression and prostaglandin E2 (PGE2) secretion, indicating that they may be independently regulated. CD147 potentially represents a novel therapeutic target for the treatment of feline OSCC and further study of CD147 is warranted. Full article

Influenza viruses are among the major infectious disease threats of animal and human health. This review examines the recent discovery of novel influenza viruses in bats and cattle, the evolving complexity of influenza virus host range including the ability to cross species barriers and geographic boundaries, and implications to animal and human health. Full article

The incidence of canine mammary carcinoma varies with age, breed, and spay status, being among the main tumors appearing in intact female dogs. Thirty-six canine mammary carcinoma patients received injections of canine interferon-β (cIFN-β) and HSV-thymidine kinase/ganciclovir (HSV-tk/GCV) carrying lipoplexes, into the tumor bed, immediately after surgery. Next, they started periodic subcutaneous injections of lipoplexes carrying a human granulocyte-macrophage colony stimulating factor and interleukin-2 mixed with allogeneic mammary carcinoma extracts. This combined strategy was safe and well tolerated. In addition, only two out of 26 patients treated with complete surgery developed a local relapse, and 0 out of 29 stage II and III patients displayed distant metastases, suggesting both local and systemic antitumor activities. The most encouraging result was the long survival times: 22 > 1 year (where 13 > 2 and 4 > 3 years), while maintaining a good quality of life. The preliminary results in five patients presenting with local disease, an additional HSV-tk/GCV plus cIFN-β gene treatment induced local antitumor activity, evidenced by four objective responses (one complete, three partial) and one stable disease. This successful outcome supports further studies to validate this approach not only for canine veterinary patients, but also for translation to human patients. Full article

Many viruses have been associated with runting and stunting syndrome (RSS). These viral infections mainly affect young chickens, causing apathy, depression, ruffled feathers, cloacal pasting, and diarrhea. Chicken Parvovirus (ChPV) is such an infection and has been detected in chickens showing signs of enteric diseases worldwide. Therefore, the present study aims to develop a sensitive real-time fast-qPCR assay based on SYBR^® Green for detection and quantification of ChPV. A 561-bp non-structural (NS) gene was amplified and cloned, and a pair of primers was designed based on conserved nucleotide sequences on the NS gene of ChPV, the intercalating DNA reagent SYBR^® Green was employed, and the Fast mode of a thermocycler was used. The assay detects 10⁹ to 10¹ copies of the genome (CG). The limit of detection (LoD) was estimated to five CG, and the limit of quantification (LoQ) was estimated at ten CG. The standard curve efficiency was 101.94%, and the melting curve showed a unique clean peak and a melting temperature of 79.3 °C. The assay was specific to amplify the ChPV NS gene, and no amplification was shown from other viral genomes or in the negative controls. A total of 141 samples were tested using the assay, of which 139 samples were found positive. The highest CG value of ChPV was 5.7 × 10⁶ CG/uL of DNA without apparent clinical signs of enteric disturbance, and 4.6 × 10⁶ CG/uL DNA were detected in chickens with RSS. Full article

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Rabies is a neglected but preventable viral zoonosis that poses a substantial threat to public health. In this regard, a global program has been initiated for the elimination of human rabies caused by rabid dogs through the mass vaccination of canine populations. Geographic areas vary greatly towards attainment of this objective. For example, while dog-mediated and wildlife rabies have been largely controlled in major parts of the Americas and Western Europe, the Middle East still grapples with human rabies transmitted by unvaccinated dogs and cats. Rabies prevention and control in the Middle East is quite difficult because the region is transcontinental, encompassing portions of Africa, Asia, and Europe, while consisting of politically, culturally, and economically diverse countries that are often subject to war and unrest. Consequently, one over-riding dilemma is the misinformation or complete lack of rabies surveillance data from this area. This communication is an attempt to provide an overview of rabies in the Middle East, as a cohesive approach for the honing of disease management in each area, based on data compiled from multiple sources. In addition, the related regional transboundary movement of rabies was investigated through phylogenetic studies of available viral gene sequences. Thereafter, the epidemiological status of rabies was assessed for the region. Finally, localities were classified first by the Stepwise Approach towards Rabies Elimination framework and then categorized into four different groups based on management theme: “rabies free”; owned dog and domestic animal vaccination; community dog vaccination; and wildlife vaccination. The classification system proposed herein may serve as a baseline for future efforts. This is especially important due to the severe lack of rabies information available for the Middle East as a whole and a need for a comprehensive program focusing on the entirety of the region in light of renewed international commitment towards canine rabies elimination. Full article

Veterinarians are in a strong position of social influence on animal-related issues. Hence, veterinary schools have an opportunity to raise animal health and welfare standards by improving veterinary students’ animal welfare and ethics (AWE) education, including that related to animals used for scientific purposes. A survey of 818 students in the early, mid, and senior stages of their courses at all eight veterinary schools across Australia and New Zealand was undertaken on their first day of practice (or Day One Competences) to explore how veterinary students viewed the importance of their competence in the management of welfare and ethical decision-making relating to animals kept for scientific purposes. From highest to lowest, the rankings they assigned were: Animal Ethics Committee (AEC) Procedures or Requirements; 3Rs (Replacement, Refinement and Reduction); Humane Endpoints; Euthanasia; “What Is a Research Animal?”; and Conscientious Objections. Female students rated Conscientious Objections, Humane Endpoints, and Euthanasia significantly higher than male students did across the three stages of study. The score patterns for these three variates showed a trend for the male students to be more likely to score these topics as extremely important as they advanced through the course, but female students’ scores tended to decline slightly or stay relatively stable. No gender differences emerged for the three variates: 3Rs (Replacement, Refinement and Reduction); AEC Procedures or Requirements; and “What Is a Research Animal?”. This study demonstrates that understandings of the regulatory and normative frameworks are considered most important in animal welfare and ethics competence in veterinary students. To the authors’ knowledge, this is the first study to investigate what importance veterinary students place on their competence regarding animals kept for scientific purposes. Full article

The importance of animal welfare and ethics (AWE) within the veterinary education should reflect community concerns and expectations about AWE, and the professional demands of veterinary accreditation on the first day of practice (or ‘Day One’ competences). Currently, much interest and debate surrounds the treatment of production animals, particularly around live export. To explore the attitudes to AWE of veterinary students in Australia and New Zealand, a survey was undertaken to (i) understand what students consider important AWE topics for initial production animal competence; and (ii) ascertain how these priorities correlated with gender, area of intended practice and stage-of-study. The results from 575 veterinary students showed that all students ranked strategies to address painful husbandry procedures as the most important issues on their first day in production animal practice. Additionally, it was found that the importance students assigned to an understanding of human–animal interactions declined as they progressed through the veterinary course. In contrast, the importance of an understanding of euthanasia issues for production animals increased for male students as they progressed through the course, and remained consistently high in females. Females also gave higher ranking to the importance of understanding production animal stress associated with transport, and ranked strategies to address painful husbandry procedures more important than did males. These findings should help the development of AWE teaching resources that address students’ attitudes and competence and that can be delivered when students are most receptive. Full article

A six-year-old female goat was presented to the veterinary teaching hospital of the University of the West Indies with a history of progressive hind-limb paresis lasting two weeks. The doe developed a grade 6/6 holosystolic murmur during hospitalisation. Echocardiography revealed vegetative growths attached to cusps of the mitral and aortic valves. There was an accelerated aortic flow at 2.9 m/s and aortic insufficiency. The aortic vegetation was prolapsing into the left ventricle during diastole, causing it to contact the septal mitral valve leaflet. A diagnosis of mitral and aortic vegetative endocarditis, with a mitral kissing vegetation and mild aortic stenosis, was reached. The patient was placed on broad-spectrum antimicrobials. A short-term follow-up showed no resolution of clinical signs, and the animal eventually died. Post-mortem examination showed severe vegetative, fibrino-necrotic, aortic and mitral valve lesions. The goat also had a severe fibrino-suppurative mastitis. Histopathology confirmed the lesions to be vegetative endocarditis. Full article

Adverse food reactions (AFR) are a common cause of skin diseases in cats and dogs. The correct diagnosis and management of AFR relies upon clinical nutrition. The reliability of commercial hypoallergenic diets commonly used in AFR has been questioned because studies have shown the presence of proteins not declared on the label ingredients. It is proposed that extensively hydrolysed protein-based diets constitute a reliable nutritional solution. Royal Canin Anallergenic™ Canine and Feline diets are formulated with very low molecular weight feather protein and purified corn starch. Protein gel electrophoresis and thin layer paper chromatography were used to characterize protein hydrolysis in these diets and their hydrolysed raw materials; protein species were identified by mass spectrometry. To detect cross-contaminating protein, species-specific DNA was measured and correlated with ancillary protein content using calibration curves. The only protein components detected in the extensively hydrolysed feather protein raw material were amino acids and small oligopeptides. GBSS-I (Granule-bound starch synthase 1) was detected in the finished diets; this has not been reported as a clinically apparent allergen in dogs or cats. The DNA threshold corresponding to the maximum acceptable level of ancillary protein was not exceeded in 99.9% of more than 2150 product batches tested and no products were released to the market with cross-contaminating proteins. These results demonstrate the extensive level of protein hydrolysis in Royal Canin Anallergenic™ Canine and Feline diets and the absence of cross-contaminating protein, both key requirements for a diet to be used during diagnosis and for management of pets with AFR. Full article

Escherichia coli and Aspergillus fumigatus are two pathogens largely present among poultry. They can cause mild or severe forms of disease, and are associated with significant economic losses. The aim of the present study was to investigate the chemical composition and the in vitro antimicrobial activity of sixteen essential oils (EOs) and five mixtures against E. coli and A. fumigatus strains previously isolated from poultry. The study was performed with the following EOs: Aloysiatryphilla, Boswelliasacra, Cinnamomum zeylanicum, Citrus aurantium, Citrus bergamia, Citrus limon, Citrus reticulata, Cymbopogon citratus, Eucalyptus globulus, Lavandula hybrida, Litsea cubeba, Ocimum basilicum, Melaleuca alternifolia, Mentha piperita, Pelargoniumgraveolens, and Syzygium aromaticum. Moreover, the following mixtures were also tested: L. cubeba and C. citratus (M1), L. cubeba and A. triphylla (M2), A. triphylla and C. citratus (M3), A. triphylla, C.citratus and L. cubeba (M4), S. aromaticum and C. zeylanicum (M5). One hundred and ninety-one compounds were identified in the tested EOs and mixtures. MIC determination found good anti-E. coli activity with C. zeylanicum (2.52 mg/mL), C. citratus (1.118 mg/mL), L. cubeba (1.106 mg/mL), M. piperita (1.14 mg/mL) and S. aromaticum (1.318 mg/mL) EOs. Among the mixtures, M5 showed the best result with a MIC value of 2.578 mg/mL. The best antimycotic activity was showed by A. triphylla (0.855 mg/mL), followed by C. citratus (0.895 mg/mL), while C. aurantium, M. piperita, B. sacra and P. graveolens did not yield any antifungal effect at the highest dilution. The mixtures exhibited no antifungal activity at all. This study shows promising results in order to use EOs in the environment for disinfection purposes in poultry farms and/or in hatcheries. Full article

Cleft palate in dogs is a congenital defect that mostly leads to euthanasia of the affected puppy. If an attempt is made to raise the puppy, it is generally fed via an orogastric tube. Here, we describe the management of cleft palate in three puppies (two Boxers, one Collie) using a customised temporary prosthesis, which allowed the puppies to be bottle-fed and successfully raised by their owners (Cases 2 and 3) and the author (Case 1). The temporary palatal prosthesis was manufactured from a mouthguard intended for human children, which is made of thermoplastic silicone. The preparation procedure was simple and cost-effective. All puppies underwent corrective surgery at 5–6 months of age. After surgery, one of the Boxer puppies showed mandibular mesioclusion, while the other two showed no aberrations. All puppies gained the same amount of weight as their littermates, although the weight gain of the two Boxers was slower than that of their littermates. In summary, this case report describes an easy and effective way to raise puppies with cleft palate until corrective surgery can be performed. Full article