Context-Related Variation in the Vocal Growling Behaviour of the Domestic Dog (Canis familiaris)

We investigated whether the growls of domestic dogs, Canis familiaris , exhibit predictable variations in a range of key acoustic parameters when they are given in two contrasting experimental contexts, and whether humans are sensitive to such variation. In a standardized experimental paradigm, an experimenter visited 32 domestic dogs and generated an aggression context and a play context. In these contexts, 204 isolated growls were recorded and subsequently acoustically analysed. Contrary to previous findings on barks, fundamental and formant frequencies of growls did not vary between the two contexts. However, growls from the aggression context were significantly longer than growls from the play context. Additionally, the temporal structure of the vocal sequences containing growls differed significantly between the contexts. In a series of psychoacoustic experiments, human listeners were not able to discriminate between growls recorded in the two contexts, however both aggressiveness and playfulness ratings were strongly influenced by dog weight, with larger dogs being perceived as more aggressive than smaller dogs. However, participants were able to attribute aggressiveness and playfulness to synthesized sequences in which growls occurred at a two rates, typical of either the aggression or the play context. We conclude that context-related variation in growling behaviour resides in the temporal structure, rather than in the acoustic composition of isolated growls, and furthermore that human listeners are able to attribute context on the basis of growling rate. In addition, human appear to have an intuitive acoustic bias toward perceiving larger dogs as more aggressive.     

Production of Haploid Zebrafish Embryos by In Vitro Fertilization

The zebrafish has become a mainstream vertebrate model that is relevant for many disciplines of scientific study. Zebrafish are especially well suited for forward genetic analysis of developmental processes due to their external fertilization, embryonic size, rapid ontogeny, and optical clarity – a constellation of traits that enable the direct observation of events ranging from gastrulation to organogenesis with a basic stereomicroscope. Further, zebrafish embryos can survive for several days in the haploid state. The production of haploid embryos in vitro is a powerful tool for mutational analysis, as it enables the identification of recessive mutant alleles present in first generation (F1) female carriers following mutagenesis in the parental (P) generation. This approach eliminates the necessity to raise multiple generations (F2, F3, etc.) which involves breeding of mutant families, thus saving the researcher time along with reducing the needs for zebrafish colony space, labor, and the husbandry costs. Although zebrafish have been used to conduct forward screens for the past several decades, there has been a steady expansion of transgenic and genome editing tools. These tools now offer a plethora of ways to create nuanced assays for next generation screens that can be used to further dissect the gene regulatory networks that drive vertebrate ontogeny. Here, we describe how to prepare haploid zebrafish embryos. This protocol can be implemented for novel future haploid screens, such as in enhancer and suppressor screens, to address the mechanisms of development for a broad number of processes and tissues that form during early embryonic stages.     

Simple Mechanisms Can Explain Social Learning in Domestic Dogs (Canis familiaris)

Recent studies have suggested that domestic dogs (Canis familiaris) engage in highly complex forms of social learning. Here, we critically assess the potential mechanisms underlying social learning in dogs using two problem‐solving tasks. In a classical detour task, the test dogs benefited from observing a demonstrator walking around a fence to obtain a reward. However, even inexperienced dogs did not show a preference for passing the fence at the same end as the demonstrator. Furthermore, dogs did not need to observe a complete demonstration by a human demonstrator to pass the task. Instead, they were just as successful in solving the problem after seeing a partial demonstration by an object passing by at the end of the fence. In contrast to earlier findings, our results suggest that stimulus enhancement (or affordance learning) might be a powerful social learning mechanism used by dogs to solve such detour problems. In the second task, we examined whether naïve dogs copy actions to solve an instrumental problem. After controlling for stimulus enhancement and other forms of social influence (e.g. social facilitation and observational conditioning), we found that dogs’ problem solving was not influenced by witnessing a skilful demonstrator (either an unknown human, a conspecific or the dog’s owner). Together, these results add to evidence suggesting that social learning may often be explained by relatively simple (but powerful) mechanisms.     

Two-item sentence comprehension by a dog (Canis familiaris)

Syntax use by non-human animals remains a controversial issue. We present here evidence that a dog may respond to verbal requests composed of two independent terms, one referring to an object and the other to an action to be performed relative to the object. A female mongrel dog, Sofia, was initially trained to respond to action (point and fetch) and object (ball, key, stick, bottle and bear) terms which were then presented as simultaneous, combinatorial requests (e.g. ball fetch, stick point). Sofia successfully responded to object-action requests presented as single sentences, and was able to flexibly generalize her performance across different contexts. These results provide empirical evidence that dogs are able to extract the information contained in complex messages and to integrate it in directed performance, an ability which is shared with other linguistically trained animals and may represent a forerunner of syntactic functioning.     

The mechanics of jumping by a dog (Canis familiaris)

A force platform has been used to obtain records of the forces exerted on the ground by an Alsatian dog, during take-off for running long jumps and standing scale jumps. The records have been analysed in conjunction with cinematograph film, taken simultaneously, and anatomical data. Stresses in the principal muscles of the hind limb, and in the triceps, have been calculated and the values obtained are compared with the stresses found by other investigators in isometric experiments with excised mammal muscles. Stresses in certain tendons and bones have been calculated, and the values obtained are compared with published values for the strength of tendon and bone. Evidence is presented that the gastrocnemius and plantaris muscles behave, in take-off for a jump, essentially as passive elastic bodies. Most of the elastic energy is probably stored in their tendons. A tendency for distal limb muscles to be pinnate, with much shorter fibres than proximal limb muscles, is noted and discussed.     

Isotope recycling in lactating dogs (Canis familiaris)

Isotope-based techniques for the measurement of water turnover, energy expenditure, and milk intake often assume that there is no recycling of isotopes once they have left the labeled animal. In experiments involving lactating females or their suckling offspring, there are several possible routes of isotope recycling. These include the consumption of labeled milk by offspring, the ingestion of labeled excreta, and the rebreathing of exhaled labeled CO(2) or water vapor by both mother and offspring. Isotope recycling might be especially important during lactation because the offspring are in close contact with each other and their mother for prolonged periods. We show here in 24- to 30-day-old domestic dog Canis familiaris puppies that there was no detectable transfer of (18)O or (2)H from labeled to unlabeled pups in two litters (16 pups, 8 labeled, 8 unlabeled) that were weaned early and independent of their mother. However, there was a significant transfer of both isotopes from labeled to unlabeled pups and from labeled pups to their mothers in nine equivalent nursing litters of the same age (27 labeled, 26 unlabeled pups). The increases in enrichment of isotopes in unlabeled offspring were greater than the increases in enrichment of the mothers. This indicates that maternal ingestion of offspring excreta and subsequent transfer of isotope in milk is not the sole pathway of recycling. Additional routes must also be important, such as exchange of isotope between pups on saliva-coated nipples and perhaps direct ingestion of excreta by unweaned young. Recycling is unlikely to be an important factor when determining maternal metabolic rate during peak lactation in domestic dogs. However, experiments that are designed to assess the energy demands of pups and isotope-based estimates of water turnover in offspring may need to take into account any effects of isotope recycling. In a theoretical example, removing the effects of recycling increased the measured energy expenditure in pups by up to 7% and increased the calculated elimination rates of both isotopes by up to 11.1% in (18)oxygen and 10.9% in (2)hydrogen.     

Sarcocystis caninum and Sarcocystis svanai n. spp. (Apicomplexa: Sarcocystidae) Associated with Severe Myositis and Hepatitis in the Domestic Dog (Canis familiaris)

There are several reports of Sarcocystis sarcocysts in muscles of dogs, but these species have not been named. Additionally, there are two reports of Sarcocystis neurona in dogs. Here, we propose two new names, Sarcocystis caninum, and Sarcocystis svanai for sarcocysts associated with clinical muscular sarcocystosis in four domestic dogs (Canis familiaris), one each from Montana and Colorado in the USA, and two from British Columbia, Canada. Only the sarcocyst stage was identified. Most of the sarcocysts identified were S. caninum. Sarcocysts were studied using light microscopy, transmission electron microscopy (TEM), and polymerase chain reaction. Based on collective results two new species, S. caninum and S. svanai were designated. Sarcocystis caninum and S. svanai were structurally distinct. Sarcocystis caninum sarcocysts were up to 1.2 mm long and up to 75 μm wide. By light microscopy, the sarcocyst wall was relatively thin and smooth. By TEM, the sarcocyst wall was “type 9”, 1–2 μm thick, and contained villar protrusions that lacked microtubules. Bradyzoites in sections were 7–9 μm long. Sarcocysts of S. svanai were few and were identified by TEM. Sarcocystis svanai sarcocysts were “type 1”, thin walled (< 0.5 μm), and the wall lacked villar protrusions but had tiny blebs that did not invaginate. DNA was extracted either from infected frozen muscle biopsies or formalin‐fixed paraffin‐embedded sections. Dogs were either singly infected with S. caninum or multiply co‐infected with S. caninum and S. svanai (the result of a mixed infection) based on multilocus DNA sequencing and morphology. BLASTn analysis established that the sarcocysts identified in these dogs were similar to, but not identical to Sarcocystis canis or Sarcocystis arctosi, parasites found to infect polar bears (Ursus maritimus) or brown bears (Ursus arctosi), respectively. However, the S. caninum sequence showed 100% identify over the 18S rRNA region sequenced to that of S. arctica, a parasite known to infect Arctic foxes (Vulpes lagopus).     

Nucleotide Polymorphisms in the Canine Noggin Gene and Their Distribution Among Dog (Canis lupus familiaris) Breeds

Noggin (NOG) is an important regulator for the signaling of bone morphogenetic proteins. In this study, we sequenced the complete coding sequence of the canine NOG gene and characterized the nucleotide polymorphisms. The sequence length varied from 717 to 729 bp, depending on the number of a 6-bp tandem repeat unit (GGCGCG), an insertion that has not been observed in other mammalian NOG genes investigated to date. It results in extensions of (Gly-Ala)3-5 in the putative NOG protein. To survey the distribution of these tandem repeat polymorphisms, we analyzed 126 individuals in seven dog breeds. We identified only three alleles: (GGCGCG)3, (GGCGCG)4, and (GGCGCG)5. Although the allele frequencies were remarkably different among the breeds, the three alleles were present in all seven of the breeds and did not show any deviation from Hardy-Weinberg equilibrium.     

Forces exerted on the ground by galloping dogs (Canis familiaris)

In quadrupedal galloping, one foot of each pair (the 'trailing' foot) is set down shortly before the other ('leading') foot. Alexander, Jayes & Ker (1980) argued that the trailing and leading feet should exert different patterns of force on the ground, to make the most effective use of tendon elasticity to reduce the energy cost of locomotion. We have used two force plates mounted in tandem in a playing field to obtain records of the forces exerted by individual feet of galloping dogs. We find that the patterns of force exerted by trailing and leading feet are much more similar than had been predicted.     

Molecular systematics of Mesocestoides sPP (cestoda: mesocestoididae) from domestic dogs (Canis familiaris) and coyotes (Canis latrans)

The genus Mesocestoides Vaillant, 1863 includes tapeworms of uncertain phylogenetic affinities and with poorly defined life histories. We previously documented 11 cases of peritoneal cestodiasis in dogs (Canis familiaris L.) in western North America caused by metacestodes of Mesocestoides spp. In the current study, DNA sequences were obtained from metacestodes collected from these dogs (n = 10), as well as proglottids from dogs (n = 3) and coyotes (Canis latrans Say, 1823 [n = 2]), and tetrathyridia representing laboratory isolates of M. corti (n = 3), and these data were analyzed phylogenetically. Two nuclear genetic markers, 18S ribosomal DNA and the second internal-transcribed spacer (ITS 2), were sequenced. Phylogenetic analysis of the 18S rDNA data recovered a monophyletic group composed of all samples of Mesocestoides spp., distinct from closely related outgroup taxa (Amurotaenia Akhmerov, 1941 and Tetrabothrius Rudolphi, 1819). Initial analysis of the ITS 2 data resolved 3 clades within Mesocestoides. Two proglottids from dogs formed a basal clade, a second clade was represented by tetrathyridial isolates, and a third clade included all other samples. Interpretation of these data from an apomorphy-based perspective identified 6 evolutionary lineages. We also assessed whether metacestodes from dogs (n = 4) are capable of asexual proliferation in laboratory mice. One tetrathyridial and 2 acephalic isolates from dogs proliferated asexually. Further investigation is warranted to determine which of the lineages represent distinct species and to determine the life history strategies of Mesocestoides spp.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species: XVI Dingo (Canis familiaris dingo) and dog (Canis familiaris)

As part of a programme of comparative measurements of P _d (diffusional water permeability) the RBCs (red blood cells) from dingo (Canis familiaris dingo) and greyhound dog (Canis familiaris) were studied. The morphologies of the dingo and greyhound RBCs [examined by light and SEM (scanning electron microscopy)] were found to be very similar, with regard to aspect ratio and size; the mean diameters were estimated to be the same (～7.2 μm) for both dingo and greyhound RBCs. The water diffusional permeability was monitored by using an Mn²⁺‐doping ¹H NMR technique at 400 MHz. The P _d (cm/s) values of dingo and greyhound RBCs were similar: 6.5×10^?3 at 25°C, 7.5×10^?3 at 30°C, 10×10^?3 at 37°C and 11.5×10^?3 at 42°C. The inhibitory effect of a mercury‐containing SH (sulfhydryl)‐modifying reagent PCMBS (p‐chloromercuribenzene sulfonate) was investigated. The maximal inhibition of dingo and greyhound RBCs was reached in 15–30 min at 37°C with 2 mmol/l PCMBS. The values of maximal inhibition were in the range 72–74% when measured at 25°C and 30°C, and ～66% at 37°C. The lowest value of P _d (corresponding to the basal permeability to water) was ～2–3×10^?3 cm/s in the temperature range 25–37°C. The E _a,d (activation energy of water diffusion) was 25 kJ/mol for dingo RBC and 23 kJ/mol for greyhound RBCs. After incubation with PCMBS, the values of E _a,d increased, reaching 46–48 kJ/mol in the condition of maximal inhibition of water exchange. The electrophoretograms of membrane polypeptides of the dingo and greyhound RBCs were compared and seen to be very similar. We postulate that the RBC parameters reported in the present study are characteristic of all canine species and, in particular in the two cases presented here, these parameters have not been changed by the peculiar Australian habitat over the millennia (as in the case of the dingo) or over shorter time periods, decades or centuries (as in the case of the domestic greyhound).     

Peptidergic regulation of feeding in the dog (Canis familiaris)

Although the incidence of obesity in the domesticated dog is high, few studies have investigated the regulation of food intake in this species. In the present study we investigated the response of the dog to a number of putative satiety agents including cholecystokinin (CCK), bombesin, calcitonin and naloxone. CCK significantly suppressed food intake during a scheduled fifteen minute meal in intact dogs and in dogs receiving total subdiaphragmatic vagotomies. Emesis occurred following injection of higher doses of CCK in most dogs. Bombesin and calcitonin reduced intake in both normal and vagotomized dogs, although higher doses of calcitonin were needed to significantly suppress feeding in vagotomized dogs compared with intact animals. Naloxone reduced feeding by as much as 60% in intact and vagotomized animals. Glucagon suppressed feeding in intact dogs, but not in vagotomized animals. Somatostatin and pancreatic polypeptide did not alter food intake. Thus the domesticated dog responds somewhat differently to some neuropeptides compared with the laboratory rat stressing the importance of examining the regulation of food intake across species.     

A Comparison of the Sensory Development of Wolves (Canis lupus lupus) and Dogs (Canis lupus familiaris)

Little is known about the development of the sensory systems of wolves. The timing of sensory development in wolves is usually extrapolated from studies on dogs, since they are members of the same species. However, early developmental differences between these two subspecies have already been identified. For example, wolves tend to approach and investigate objects in their environment 2 wk before dogs. These changes in developmental timing may play an important role in the behavioral differences between adult wolves and dogs. The purpose of this study is to compare the development of the sensory systems in wolves and dogs. Responses of seven wolf pups and 43 dog pups to familiar and novel olfactory, auditory, and visual stimuli were tested weekly from 2–7 wk of age. Eleven wolf pups were also observed for orientation towards auditory and visual stimuli during 2‐h sessions, 5 d a week, from 2–8 wk of age. These observations were supplemented by the daily records of caretakers. The results suggest that wolves and dogs both develop olfaction by 2 wk, audition by 4 wk, and vision by 6 wk on average, despite the 2‐wk shift in their ability to explore. This means that when wolves begin to explore at 2 wk, they are blind and deaf, and must rely primarily on their sense of smell. Thus, there is a significant alteration of how these subspecies experience their environment during the critical period of socialization. These findings lead to an alternative explanation for the difference in dogs' and wolves' abilities to form interspecies social attachments, such as those with humans.     

On the allometry of long bones in dogs (Canis familiaris)

The allometric relations of diameter and length of humerus, ulna, femur, and tibia of 108 specimens, from 63 different breeds of dogs and 12 specimens of wolves, were calculated by means of model II of regression or major axis method. Only for the tibia were the values of wolves included in the cluster formed for dog breeds. Consequently, separate lines of regression were calculated for the other bones. Results agree in general with the exponents predicted by the theory of geometric similarity; however, the slope obtained for femur (0.865) differed significantly from this. Morphology of the long bones of the legs does not differentiate dogs and wolves; this probably reflects secondary convergence among wolves with relatively modern breeds of dogs.     

Morphological and Gene Expression Changes in Cattle Embryos from Hatched Blastocyst to Early Gastrulation Stages after Transfer of In Vitro Produced Embryos

A detailed morphological staging system for cattle embryos at stages following blastocyst hatching and preceding gastrulation is presented here together with spatiotemporal mapping of gene expression for BMP4, BRACHYURY, CERBERUS1 (CER1), CRIPTO, EOMESODERMIN, FURIN and NODAL. Five stages are defined based on distinct developmental events. The first of these is the differentiation of the visceral hypoblast underlying the epiblast, from the parietal hypoblast underlying the mural trophoblast. The second concerns the formation of an asymmetrically positioned, morphologically recognisable region within the visceral hypoblast that is marked by the presence of CER1 and absence of BMP4 expression. We have termed this the anterior visceral hypoblast or AVH. Intra-epiblast cavity formation and the disappearance of the polar trophoblast overlying the epiblast (Rauber’s layer) have been mapped in relation to AVH formation. The third chronological event involves the transition of the epiblast into the embryonic ectoderm with concomitant onset of posterior NODAL, EOMES and BRACHYURY expression. Lastly, gastrulation commences as the posterior medial embryonic ectoderm layer thickens to form the primitive streak and cells ingress between the embryonic ectoderm and hypoblast. At this stage a novel domain of CER1 expression is seen whereas the AVH disappears. Comparison with the mouse reveals that while gene expression patterns at the onset of gastrulation are well conserved, asymmetry establishment, which relies on extraembryonic tissues such as the hypoblast and trophoblast, has diverged in terms of both gene expression and morphology.     

Robertsonian translocation (8;14) in an infertile bitch (Canis familiaris)

A new case of a Robertsonian translocation diagnosed in an infertile bitch of West Highland White Terrier breed is reported. Cytogenetic analyses were performed with the use of Giemsa staining, Q-, C- and Ag-I banding techniques and FISH with canine whole X chromosome paint. The karyotype of this bitch was described as 77,XX der (8;14)(q10;q10).     

Mechanics of locomotion of dogs (Canis familiaris) and sheep (Ovis aries)

Records have been made of the forces exerted on the ground by dogs and a sheep, in walking, trotting, cantering and slow galloping. Film has been taken simultaneously. The difference between walking and trotting was much less marked for the sheep than for the dogs.
Step length and stride length increase as speed increases. They are expressed as functions of the Froude number.
The vertical component of the force exerted by a foot on the ground shows two main maxima in walking, except in the case of the fore feet of sheep. In this case and in other gaits there is only one main maximum. The vertical movements of the fore and hind quarters which occurred in examples of each gait have been calculated from the force records.
The force exerted by a foot on the ground changes direction in the course of a step so as to remain more or less in line with a point fixed relative to the animal, but dorsal to its back.
The force records show impact disturbances in the first 003 sec of contact of each foot with the ground.
The point of application of the force on the sole of a foot tends to move posteriorly as the force increases.
The results are discussed in relation to a theoretical account of the mechanics of locomotion on legs.