Structural basis and analysis of hamster ACE2 binding to different SARS-CoV-2 spike RBDs.

Pet golden hamsters were first identified being infected with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) delta variant of concern (VOC) and transmitted the virus back to humans in Hong Kong in January 2022. Here, we studied the binding of two hamster (golden hamster and Chinese hamster) angiotensin-converting enzyme 2 (ACE2) proteins to the spike protein receptor-binding domains (RBDs) of SARS-CoV-2 prototype and eight variants, including alpha, beta, gamma, delta, and four omicron sub-variants (BA.1, BA.2, BA.3, and BA.4/BA.5). We found that the two hamster ACE2s present slightly lower affinity for the RBDs of all nine SARS-CoV-2 viruses tested than human ACE2 (hACE2). Furthermore, the similar infectivity to host cells expressing hamster ACE2s and hACE2 was confirmed with the nine pseudotyped SARS-CoV-2 viruses. Additionally, we determined two cryo-electron microscopy (EM) complex structures of golden hamster ACE2 (ghACE2)/delta RBD and ghACE2/omicron BA.3 RBD. The residues Q34 and N82, which exist in many rodent ACE2s, are responsible for the lower binding affinity of ghACE2 compared to hACE2. These findings suggest that all SARS-CoV-2 VOCs may infect hamsters, highlighting the necessity of further surveillance of SARS-CoV-2 in these animals.IMPORTANCESARS-CoV-2 can infect many domestic animals, including hamsters. There is an urgent need to understand the binding mechanism of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants to hamster receptors. Herein, we showed that two hamster angiotensin-converting enzyme 2s (ACE2s) (golden hamster ACE2 and Chinese hamster ACE2) can bind to the spike protein receptor-binding domains (RBDs) of SARS-CoV-2 prototype and eight variants and that pseudotyped SARS-CoV-2 viruses can infect hamster ACE2-expressing cells. The binding pattern of golden hamster ACE2 to SARS-CoV-2 RBDs is similar to that of Chinese hamster ACE2. The two hamster ACE2s present slightly lower affinity for the RBDs of all nine SARS-CoV-2 viruses tested than human ACE2. We solved the cryo-electron microscopy (EM) structures of golden hamster ACE2 in complex with delta RBD and omicron BA.3 RBD and found that residues Q34 and N82 are responsible for the lower binding affinity of ghACE2 compared to hACE2. Our work provides valuable information for understanding the cross-species transmission mechanism of SARS-CoV-2.

The impact of state cannabis legislation, county-level socioeconomic and dog-level characteristics on reported cannabis poisonings of companion dogs in the USA (2009-2014).

With current trends in cannabis legalization, large efforts are being made to understand the effects of less restricted legislation on human consumption, health, and abuse of these products. Little is known about the effects of cannabis legalization and increased cannabis use on vulnerable populations, such as dogs. The objective of this study was to examine the effects of different state-level cannabis legislation, county-level socioeconomic factors, and dog-level characteristics on dog cannabis poisoning reports to an animal poison control center (APCC). Data were obtained concerning reports of dog poisoning events, county characteristics, and state cannabis legislation from the American Society for the Prevention of Cruelty to Animals' (ASPCA) APCC, the US Census Bureau, and various public policy-oriented and government websites, respectively. A multilevel logistic regression model with random intercepts for county and state was fitted to investigate the associations between the odds of a call to the APCC being related to a dog being poisoned by a cannabis product and the following types of variables: dog characteristics, county-level socioeconomic characteristics, and the type of state-level cannabis legislation. There were significantly higher odds of a call being related to cannabis in states with lower penalties for cannabis use and possession. The odds of these calls were higher in counties with higher income variability, higher percentage of urban population, and among smaller, male, and intact dogs. These calls increased throughout the study period (2009-2014). Reporting of cannabis poisonings were more likely to come from veterinarians than dog owners. Reported dog poisonings due to cannabis appear to be influenced by dog-level and community-level factors. This study may increase awareness to the public, public health, and veterinary communities of the effects of recreational drug use on dog populations. This study highlights the need to educate dog owners about safeguarding cannabis products from vulnerable populations.

Comparison of methodologies for fat determination and evaluation of calcium and phosphorus content in snacks for dogs / Comparação de metodologias para determinação de gordura e avaliação do conteúdo de cálcio e fósforo em petiscos para cães

In recent years, the relationship between humans and companion animals has tightened considerably and resulted in the expansion of the range of pet food industry products available in the market. In this context, snacks have gained greater popularity as pet owners seek to please their animals by providing such foods. Due to the growing importance of the snack segment, a need exists for accurate information on the nutritional composition of these products, such as fat concentration. No studies were found that evaluated the effectiveness of different methods applied for determining the content of this nutrient in dog snacks. In addition, too much mineral content can pose health risks. Thus, the objective of this study was to compare three methodologies for determining fat in pet snack products. The moisture, calcium and phosphorus content of each was also determined to compare the obtained results with each value stated on their product labels. Fat determination methods evaluated were ether extract (EE), ether extract after acid hydrolysis (EEHA), and fat content obtained from Ankom XT15 analyzer (ANKOM). Twenty-four snacks produced by 17 companies were evaluated. The results of the three methodologies were compared using the Tukey test. The comparison between the results of the laboratory analysis and the values stated on the labels was performed using descriptive statistics. There was no difference between the three methods evaluated (p = 0.34) regarding fat content. Regarding the nutritional compliance of the labels, 25% (n = 6) of the snacks presented higher moisture content than the declared amount, 50% (n = 12) presented lower fat content, 25% (n = 6) lower phosphorus content and, in 50% (n = 12), the calcium content was not within the minimum and maximum range stated on the label. Therefore, due to the absence of difference between the results, any of the three fat determination methodologies could be used. Regarding compliance of labels for calcium, phosphorus and fat content, greater control over the nutritional composition of these foods is required since most pet owners tend to supply large quantities of snacks to dogs, leading to excessive daily energy intake.(AU)

Nos últimos anos a relação entre seres humanos e animais de companhia estreitou-se consideravelmente e houve uma expansão da gama de produtos da indústria pet food disponíveis no mercado. Nesse contexto, os petiscos ganharam maior popularidade, uma vez que os tutores buscam agradar seus pets com esse tipo de alimento. Devido à crescente importância do segmento de petiscos, há a necessidade de informações precisas sobre a composição nutricional desses produtos, como o teor de gordura, uma vez que não foram encontrados estudos que avaliassem a eficácia dos métodos de determinação do teor deste nutriente em petiscos destinados a cães e o excesso de minerais pode implicar em riscos para a saúde. Assim, o presente trabalho comparou três metodologias para determinação de gordura em petiscos para cães, e também determinou os seus respectivos teores de umidade, cálcio e fósforo, cujos resultados foram comparados aos valores declarados pelos fabricantes nos rótulos dos produtos. Os métodos de determinação da gordura avaliados foram: extrato etéreo (EE), extrato etéreo após hidrólise ácida (EEHA) e teor de gordura obtido em analisador Ankom XT15 (ANKOM). Vinte e quatro petiscos produzidos por 17 empresas foram avaliados. Os resultados das três metodologias de determinação da gordura foram comparados com o emprego do teste Tukey. A comparação entre os resultados das análises laboratoriais e os valores declarados nos rótulos foi realizada por meio de estatística descritiva. Não houve diferença entre os três métodos avaliados (p = 0,34) em relação ao teor de gordura dos petiscos examinados. Em relação à conformidade nutricional dos rótulos, 25% (n = 6) dos petiscos apresentaram teor de umidade superior ao declarado, 50% (n = 12) apresentaram menor teor de gordura; 25% (n = 6) menor teor de fósforo e, em 50% (n = 12) deles, o teor de cálcio estava fora da faixa mínima e máxima declarada no rótulo. Portanto, devido à ausência de diferença entre os resultados, as três metodologias de determinação de gordura podem ser utilizadas. Quanto à conformidade dos rótulos em relação aos teores de cálcio, fósforo e gordura, é necessário maior controle sobre a composição nutricional desses alimentos, uma vez que a maioria dos tutores fornece petiscos em elevadas quantidades para os cães, que podem determinar excessivo consumo de energia.(AU)

Dietary supplementation with flaxseed mucilage alone or in combination with calcium in dogs: effects on apparent digestibility of fat and energy and fecal characteristics.

BACKGROUND: In humans, dietary supplementation with flaxseed mucilage and calcium decrease apparent digestibility of fat and energy. These supplements could prove useful for weight management in dogs. OBJECTIVE: To examine dry matter, energy and fat apparent digestibility, and fecal characteristics following dietary flaxseed mucilage supplementation alone or in combination with calcium. METHODS: A single-blinded crossover feeding trial was conducted on 11 privately owned dogs. During three consecutive 14-day periods, dogs where fed commercial dog food supplemented with potato starch (control diet), flaxseed mucilage or flaxseed mucilage and calcium. Feces from the past 2 days of each period were collected for analysis. Owners recorded fecal score (1-7: 1=very hard/dry feces, 2-3=ideal and 7=diarrhea). RESULTS: Apparent digestibility of fat was lower in both flaxseed mucilage diet (94.5±0.8%), and flaxseed mucilage and calcium diet (92.9±0.9%) compared with control diet (96.9±0.2%, P<0.0001) with fat digestibility in flaxseed mucilage and calcium diet being significantly lower than the diet supplemented with only flaxseed mucilage. Dry matter and energy digestibility was not significantly affected by diet. Fecal wet weight, dry weight and dry matter percentage was not affected by diet despite a higher fecal score for test diets (3.7±0.3) compared with control (2.8±0.2, P<0.007). CONCLUSION: In dogs, flaxseed mucilage decreased fat apparent digestibility and this effect was enhanced when combined with calcium. Dry matter and energy apparent digestibility was not affected. Decreased fecal quality may limit the acceptable level of supplementation. Further studies on incorporating flaxseed mucilage in pet food products for weight management are needed.

Widespread recombination, reassortment, and transmission of unbalanced compound viral genotypes in natural arenavirus infections.

Arenaviruses are one of the largest families of human hemorrhagic fever viruses and are known to infect both mammals and snakes. Arenaviruses package a large (L) and small (S) genome segment in their virions. For segmented RNA viruses like these, novel genotypes can be generated through mutation, recombination, and reassortment. Although it is believed that an ancient recombination event led to the emergence of a new lineage of mammalian arenaviruses, neither recombination nor reassortment has been definitively documented in natural arenavirus infections. Here, we used metagenomic sequencing to survey the viral diversity present in captive arenavirus-infected snakes. From 48 infected animals, we determined the complete or near complete sequence of 210 genome segments that grouped into 23 L and 11 S genotypes. The majority of snakes were multiply infected, with up to 4 distinct S and 11 distinct L segment genotypes in individual animals. This S/L imbalance was typical: in all cases intrahost L segment genotypes outnumbered S genotypes, and a particular S segment genotype dominated in individual animals and at a population level. We corroborated sequencing results by qRT-PCR and virus isolation, and isolates replicated as ensembles in culture. Numerous instances of recombination and reassortment were detected, including recombinant segments with unusual organizations featuring 2 intergenic regions and superfluous content, which were capable of stable replication and transmission despite their atypical structures. Overall, this represents intrahost diversity of an extent and form that goes well beyond what has been observed for arenaviruses or for viruses in general. This diversity can be plausibly attributed to the captive intermingling of sub-clinically infected wild-caught snakes. Thus, beyond providing a unique opportunity to study arenavirus evolution and adaptation, these findings allow the investigation of unintended anthropogenic impacts on viral ecology, diversity, and disease potential.

An emerging public health problem: acquired carbapenemase-producing microorganisms are present in food-producing animals, their environment, companion animals and wild birds.

Worldwide, the emergence and global spread of microorganisms with acquired carbapenemases is of great concern. The reservoirs for such organisms are increasing, not only in hospitals, but also in the community and environment. A new and important development is the presence of such organisms in livestock, companion animals and wildlife. During the last three years, carbapenemase-producing Escherichia coli, Salmonella spp. (VIM-1 producers) and Acinetobacter spp. (producing OXA-23 and NDM-1) in livestock animals (poultry, cattle and swine) and their environment have been reported. In addition, the isolation of NDM-1-producing E. coli, OXA-48 in E. coli and Klebsiella pneumoniae or OXA-23 in Acinetobacter spp. from companion animals (cats, dogs or horses) has also been observed. Other reports have described the presence of NDM-1-producing Salmonella isolated from wild birds, as well as OXA-23-like-producing Acinetobacter baumannii in ectoparasites. However, until now carbapenemase producers from foods have not been detected. For humans in contrast carbapenem-producing Salmonella isolates are increasingly reported. The real prevalence of carbapenemase-encoding genes in zoonotic bacteria or commensals from animals is unknown. Consequently, there is a need for intensified surveillance on the occurrence of carbapenemase-producing bacteria in the food chain and other animal sources in order to assist in the formulation of measures to prevent their potential spread.

Avian-specific real-time PCR assay for authenticity control in farm animal feeds and pet foods.

A highly sensitive TaqMan real-time PCR assay targeting the mitochondrial 12S rRNA gene was developed for detection of an avian-specific DNA fragment (68bp) in farm animal and pet feeds. The specificity of the assay was verified against a wide representation of animal and plant species. Applicability assessment of the avian real-time PCR was conducted through representative analysis of two types of compound feeds: industrial farm animal feeds (n=60) subjected to extreme temperatures, and commercial dog and cat feeds (n=210). Results obtained demonstrated the suitability of the real-time PCR assay to detect the presence of low percentages of highly processed avian material in the feed samples analysed. Although quantification results were well reproducible under the experimental conditions tested, an accurate estimation of the target content in feeds is impossible in practice. Nevertheless, the method may be useful as an alternative tool for traceability purposes within the framework of feed control.

Alternative dietary fiber sources in companion animal nutrition.

The US has a pet population of approximately 70 million dogs and 74 million cats. Humans have developed a strong emotional bond with companion animals. As a consequence, pet owners seek ways to improve health, quality of life and longevity of their pets. Advances in canine and feline nutrition have contributed to improved longevity and well-being. Dietary fibers have gained renewed interest in the pet food industry, due to their important role in affecting laxation and stool quality. More recently, because of increased awareness of the beneficial effects of dietary fibers in health, as well as the popularity of functional foods and holistic and natural diets, alternative and novel carbohydrates have become widespread in human and pet nutrition. Fiber sources from cereal grains, whole grains and fruits have received increasing attention by the pet food industry and pet owners. While limited scientific information is available on the nutritional and nutraceutical properties of alternative fiber sources, studies indicate that corn fiber is an efficacious fiber source for pets, showing no detrimental effects on palatability or nutrient digestibility, while lowering the glycemic response in adult dogs. Fruit fiber and pomaces have good water-binding properties, which may be advantageous in wet pet food production, where a greater water content is required, along with low water activity and a firm texture of the final product. Rice bran is a palatable fiber source for dogs and may be an economical alternative to prebiotic supplementation of pet foods. However, it increases the dietary requirement of taurine in cats. Barley up to 40% in a dry extruded diet is well tolerated by adult dogs. In addition, consumption of complex carbohydrates has shown a protective effect on cardiovascular disease and oxidative stress. Alternative fiber sources are suitable ingredients for pet foods. They have been shown to be nutritionally adequate and to have potential nutraceutical properties.

Case series: inhaled coral vapor--toxicity in a tank.

INTRODUCTION: Palytoxin (PTX) is considered a severe marine toxin. Although rare, reports of human exposure from consumption of PTX have described significant morbidity and mortality. PTX is the suspected agent in Haff disease, in which rhabdomyolysis occurs within 24 h of eating contaminated fish such as buffalo fish. PTX is primarily present in soft corals or in dinoflagellates, and it can contaminate crustaceans and other fish as it bioaccumulates up the food chain. Only 23 cases have been reported in the USA, including two recent cases in New York City. Reports of inhalational exposure to PTX are uncommon. CASE REPORTS: We describe a case series of six patients, including four adults and two children, with inhalational exposure to PTX aerosolized from Palythoa corals. Their symptoms included some degree of respiratory involvement, myalgias, paresthesias, low-grade fevers, and gastrointestinal symptoms. Fortunately, there were no serious outcomes and all patients survived without sequelae. DISCUSSION: Although rare, exposure to palytoxin is not restricted to people visiting marine environments because of Palythoa coral in some home aquariums. Routes of exposure go beyond consumption of fish that feed on the coral and include dermal as well as inhalational exposure. Palytoxin exposure should be considered in the differential diagnosis of patients who own or work with fish tanks and present with symptoms that include respiratory complaints, myalgias, neuromuscular dysfunction, hemolysis, and cardiac toxicity. There is no known antidotal therapy and treatment should focus on meticulous supportive care.