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[Development cycle of cattle sarcosporidia].

To establish the circulation of bovine sarcosporidia among wild animals of Uzbekistan foxes and young wolves were fed with bovine hearts and oesophaguses containing cysts of Sarkocystis bovicanis (= S. fusiformis). These animals were proved to be the final hosts of this species in nature. The prepatent period of S. bovicanis was found to be 7-8 and 7-9 days in foxes and young wolves, respectively and the patent one 33 and 28-34 days, respectively.

Animals

Canid modification of human remains: implications for time-since-death estimations.

Time-since-death estimations are usually based on physical decomposition of the corpse, insect succession, and contextual associations. The rates of change and succession are based on decomposition studies, most of which control access of scavengers to the corpse; however, many naturally exposed corpses are subject to scavenger modification. These modifications change the rate of decomposition, the pattern of insect succession, and the context of associations, thus altering estimations of time since death. A controlled feeding study with captive wolves and road-killed deer is pertinent to understanding canid scavenging and how scavenging may alter postmortem changes. During feeding, the wolves commonly dismember and devour the deer in a predictable sequence. Although there are some variations in the usual sequence, the carcass is always moved, and skeletal elements are separated, diminished in size and scattered. Scavenging must, therefore, be considered in estimating time since death.

Animals

Genomic signatures of the Arctic-adapted North American gray wolf ecotype.

The Arctic Circle is one of Earth's most extreme environments. It features cold temperatures, resource shortages, and near-complete winter darkness. Here, we generated a chromosome-level genome assembly of a male wolf from the Arctic Archipelago (Canis lupus arctos). Our assembly and that of the related C. l. orion from Greenland was used to identify candidate genic and regulatory features of Arctic-adapted polar wolves, ranging from selection acting on standing variation and amino acid changes in genes to conserved non-exonic elements (CNEEs) that may regulate gene expression. We identified genes and nearby CNEEs associated with thermoregulation (e.g., APOB), coat color and patterning (e.g., GOLGB1), and DNA damage response (e.g., POLQ). In vitro assays supported changes in polar wolf gene (POLQ and TRPV2 amino acid substitutions) and CNEE function. Our report offers insights into the genetic mechanisms underlying polar wolf adaptations, laying a foundation for future studies on Arctic canines.

Amino acid substitutions

Integrative Genotyping and Analysis of Canine Structural Variation Using Long-read and Short-read Data.

Structural variation makes an important contribution to canine evolution and phenotypic differences. Although recent advances in long-read sequencing have enabled the generation of multiple canine genome assemblies, most prior analyses of structural variation have relied on short-read sequencing. To offer a more complete assessment of structural variation in canines, we performed an integrative analysis of structural variants present in 12 canine samples with available long-read and short-read sequencing data along with genome assemblies. Use of long-reads permits the discovery of heterozygous variation that is absent in existing haploid assembly representations while offering a marked increase in the ability to identify insertion variants relative to short-read approaches. Examination of the size spectrum of structural variants shows that dimorphic LINE-1 and SINE variants account for over 45% of all deletions and identified 1,410 LINE-1s with intact open reading frames that show presence-absence dimorphism. Using a graph-based approach, we genotype newly discovered structural variants in an existing collection of 1,879 resequenced dogs and wolves, generating a variant catalog containing a 56.5% increase in the number of deletions and 705% increase in the number of insertions previously found in the analyzed samples. Examination of allele frequencies across admixture components present across breed clades identified 283 structural variants evolving with a signature of selection.

Animals

Topography of ganglion cells in the dog and wolf retina.

The topographical distribution of retinal ganglion cells in seven breeds of dog (Canis lupus f. familiaris) and in the wolf (Canis lupus) was studied in retinal wholemounts stained with cresyl violet or with a reduced silver method. A prominent feature of all wolf retinae was a pronounced "visual streak" of high ganglion cell density, extending from the central area far into both temporal and nasal retina. By contrast, either a pronounced or a moderate visual streak was found in dog retinae. It is hypothesized that a pronounced streak is an archetypal feature of Canis lupus, and that the moderate streak in some dogs is a corollary of breeding during domestication. Irrespective of the differences in streak form and retinal area, the estimated total number of ganglion cells was about 200,000 cells in the wolf and 115,000 in the dog. Ganglion cell density maxima in the central area of the wolf were about 12,000-14,000/mm2, and in the dog they ranged from 6,400/mm2 to 14,400/mm2. This implies individual differences in visual acuity. Alpha ganglion cells constituted 3-14% of all ganglion cells in the dog and 1-18% in the wolf, depending on retinal location. A distinct feature of all dogs and wolves was the absence of alpha cells in a substantial region of temporal peripheral retina. This has not been found in any other mammalian species and suggests corresponding functional deficits.

Animals

Catecholaminergic amacrine cells in the dog and wolf retina.

Catecholaminergic (presumed dopaminergic) amacrine cells in the retinae of Beagle dogs (canis lupus f. familiaris) and wolves (canis lupus) were visualized with an antiserum against tyrosine hydroxylase (TH). In both species, TH immunoreactivity is found in a population of amacrine cells with large somata (about 14 microns diameter) and large, moderately branched dendritic trees. Somata are located in the proximal inner nuclear layer (normal amacrines) or in the ganglion cell layer (displaced amacrines). Most dendrites stratify in a narrow band in the inner plexiform layer close to the inner nuclear layer, where they form a dense plexus with the characteristic pattern of "dendritic rings." The displaced cells have some of their dendrites in a proximal stratum of the inner plexiform layer. A few immunopositive processes are found in the outer plexiform layer (interplexiform processes). In Beagle dogs, the cell density of catecholaminergic amacrines varies from less than 1/mm2 in far periphery to 40-55/mm2 in central retina (mean density 21/mm2). The proportion of displaced amacrines varies locally from 10 to 85% (overall proportion 41% in one retina). In the wolf, densities of catecholaminergic cells range between about 3/mm2 in peripheral and up to 35/mm2 in central retina. The proportion of displaced cells is somewhat lower than in dogs, varying between 11 and 31% across the retina. The morphology and density distribution of canine catecholaminergic amacrines resemble that of other mammalian retinae. A marked difference, however, is the high percentage of displaced cells in both dog and wolf retina; it is the highest found in any mammal so far. The displaced and normal cells appear to be members of a single functional population. A comparison of the topographic distributions of catecholaminergic amacrines, rods, and ganglion cells in the dog retina shows no consistent density correlations between these neurons that are all part of the rod pathway.

Animals

Food caching in captive coyotes: stereotypy of action sequence and spatial distribution of cache sites.

This paper describes some aspects of the food caching behaviour of four captive coyotes. Detailed observations of the actions used by coyotes to cache food revealed them to be strikingly similar to those previously described for timber wolves. The similarities included the identity of the movements used, their temporal sequencing, and their susceptibility to interruption. This suggests that there exists a stereotypy across canids in the action sequences used in caching. Second, an examination of the distribution of cache sites revealed that each coyote scattered cache sites widely within a wooded region of their enclosure and preferentially in terrains close to exposed tree roots.

Animals

DNA phylogeny of the extinct marsupial wolf.

The phylogenetic affiliation of the extinct marsupial wolf (Thylacinus cynocephalus), which once was widespread in Australia, has been uncertain. On the basis of morphology, some systematists argue that the thylacine was most closely related to an extinct group of South American carnivorous marsupials, the borhyaenids, whereas others consider it to be closer to Australian carnivorous marsupials. Here we use direct sequencing by means of the polymerase chain reaction (PCR) to compare 219 bases of mitochondrial (mt) DNA from museum specimens of the marsupial wolf and representatives of six genera of extant marsupials. In agreement with the results of an antigenic study of albumin, our genetic data suggest that the marsupial wolf was more closely related to other Australian marsupial carnivores than to those of South America. Thus, the marsupial wolf represents an example of convergent morphological evolution to South American carnivorous marsupials as well as to true wolves.

Amino Acid Sequence

Growth stage dependent expression of MHC antigens on the canine transmissible venereal sarcoma.

Canine transmissible venereal sarcoma (CTVS) is a naturally occurring contagious neoplasm which can be transplanted with intact viable cells across major histocompatibility (MHC) barriers within the species and even to other members of the canine family, such as foxes, coyotes, and wolves. After 2 to 4 months of progressive growth the tumour regresses spontaneously in adults but metastasizes in immunosuppressed hosts and neonates. The mechanisms of how the tumour cells manage to overcome histocompatibility barriers so successfully for such a long period and yet succumb later are not known. In the present study we found that CTVS cells were not stimulatory to the lymphocytes of normal or tumour-bearing animals in mixed lymphocyte-tumour reaction (MLTR), although the lymphocytes from tumour-bearing animals in mixed lymphocyte-tumour reaction (MLTR), although the lymphocytes from tumour-bearing hosts responded well to either phytohaemagglutinin (PHA) or third-party allogeneic lymphocytes. Immunofluorescent antibody (IFA) assay of MHC antigens by monoclonal antibodies (MoAb) to monomorphic Class I and Class II MHC antigens showed that progressor tumour cells lacked the expression of the antigens whereas 30 to 40% of regressor tumour cells expressed them.

Animals

Skull morphology of the extinct Tasmanian tiger suggests unique biting style.

The recently extinct thylacine (Tasmanian tiger) was the largest modern marsupial predator. It is considered a classic example of evolutionary convergence due to striking similarities with placental canids (e.g., foxes and wolves), particularly in the skull, despite ~160 million years of evolutionary separation. However, we here present geometric and linear morphometric evidence that the thylacine's cranial form arises from a mosaic of traits not represented among canids or other living mammalian carnivores. Thylacines had disproportionately large heads, tall and gracile snouts with a flared canine region, and conspicuously large infraorbital foramina. Many of these traits suggest adaptations to fast, high-impact snapping behaviour in prey capture, as proposed for several living and extinct predatorial vertebrates with similar trait combinations. The thylacine's cranial function may therefore not be inferable from observation of living mammals. However, genomic progress presents new opportunities for future insights into the evolution and development of thylacine cranial adaptation.

Animals

Evolutionary consequences of domestication on the selective effects of new amino acid changing mutations in canids.

The domestication of wild canids led to dogs no longer living in the wild but instead residing alongside humans. Extreme changes in behavior and diet associated with domestication may have led to the relaxation of the selective pressure on traits that may be less important in the domesticated context. Thus, here we hypothesize that strongly deleterious mutations may have become less deleterious in domesticated populations. We test this hypothesis by estimating the distribution of fitness effects (DFE) for new amino acid changing mutations using whole-genome sequence data from 24 gray wolves and 61 breed dogs. We find that the DFE is strikingly similar across canids, with 26-28% of new amino acid changing mutations being neutral/nearly neutral (|s| < 1e-5), and 41-48% under strong purifying selection (|s| > 1e-2). Our results are robust to different model assumptions suggesting that the DFE is stable across short evolutionary timescales, even in the face of putative drastic changes in the selective pressure caused by artificial selection during domestication and breed formation. On par with previous works describing DFE evolution, our data indicate that the DFE of amino acid changing mutations depends more strongly on genome structure and organismal characteristics, and less so on shifting selective pressures or environmental factors. Given the constant DFE and previous data showing that genetic variants that differentiate wolf and dog populations are enriched in regulatory elements, we speculate that domestication may have had a larger impact on regulatory variation than on amino acid changing mutations.

Journal Article

The distribution of fitness effects varies phylogenetically across animals.

The distribution of fitness effects (DFE) describes the selection coefficients () of newly arising mutations and fundamentally influences population genetic processes. However, the extent and mechanisms of DFE variation have not been systematically investigated across species with divergent phylogenetic histories and ecological functions. Here, we inferred the DFE in natural populations of eleven animal (sub)species, including humans, mice, fin whales, vaquitas, wolves, collared flycatchers, pied flycatchers, halictid bees, Drosophila, and mosquitoes. We find that the DFE co-varies with phylogeny, where the expected mutation effects are more similar in closely related species (). Additionally, mammals have a higher proportion of strongly deleterious mutations (22% to 47% in mammals; 0.0% to 5.4% in insects and birds) and a lower proportion of weakly deleterious mutations than insects and birds. Population size is significantly negatively correlated with the expected impact of new deleterious mutations (), and the proportion of new beneficial mutations (). These findings align with Fisher's Geometric Model (FGM), which defines organismal complexity as the number of phenotypes under selection. Consistent with the FGM's predictions, we observe that mutations are more deleterious in complex organisms, while beneficial mutations occur more frequently in smaller populations to compensate for the drift load. Our study demonstrates strong phylogenetic constraints in the evolution of a fundamental population genetics parameter, and proposes that, through mechanisms of global epistasis, long-term population size and organismal complexity drive variation in the DFE across animals.

Fisher&#x2019;s geometric model

The distribution of fitness effects of nonsynonymous mutations varies phylogenetically across animals.

The distribution of fitness effects (DFE) describes the selection coefficients of newly arising mutations and fundamentally influences population genetic processes. However, the extent and mechanisms of differences in the DFE for non-synonymous mutations have not been systematically investigated across species with divergent phylogenetic histories and ecologies. Here, we inferred the DFE in natural populations of 11 animal (sub)species, including humans, mice, fin whales, vaquitas, wolves, collared flycatchers, pied flycatchers, halictid bees, Drosophila, and mosquitoes. We found that mammals have a higher proportion of strongly deleterious mutations (defined as s&#x2264;-0.01; 22% to 47% in mammals; 0.0% to 5.4% in insects and birds) and a lower proportion of weakly deleterious mutations than insects and birds. Further, the DFE co-varies with phylogeny, such that the mean mutation effects are more similar in closely related species (Pagel's &#x3bb; = 0.84, P&#x2009;=&#x2009;0.01). Next, we investigated whether various summary statistics of the DFE were related to variation in life-history traits across these organisms. We found some support for genome size, body mass, and long-term effective population size being correlated with the DFE. Overall, our findings are consistent with predictions derived independently from the Fisher's Geometric Model (FGM), which defines organismal complexity as the number of phenotypes under selection. FGM predicts that mutations are more deleterious in complex organisms, while strongly deleterious mutations occur more frequently in smaller populations. Our study demonstrates strong phylogenetic signal in the evolution of a fundamental population genetics parameter, and proposes that, through mechanisms of epistasis, long-term population size and organismal complexity could be underlying variation in the DFE across animals.

Journal Article

OCA2 splice site variant in German Spitz dogs with oculocutaneous albinism.

We investigated a German Spitz family where the mating of a black male to a white female had yielded three puppies with an unexpected light brown coat color, lightly pigmented lips and noses, and blue eyes. Combined linkage and homozygosity analysis based on a fully penetrant monogenic autosomal recessive mode of inheritance identified a critical interval of 15 Mb on chromosome 3. We obtained whole genome sequence data from one affected dog, three wolves, and 188 control dogs. Filtering for private variants revealed a single variant with predicted high impact in the critical interval in LOC100855460 (XM_005618224.1:c.377+2T>G LT844587.1:c.-45+2T>G). The variant perfectly co-segregated with the phenotype in the family. We genotyped 181 control dogs with normal pigmentation from diverse breeds including 22 unrelated German Spitz dogs, which were all homozygous wildtype. Comparative sequence analyses revealed that LOC100855460 actually represents the 5'-end of the canine OCA2 gene. The CanFam 3.1 reference genome assembly is incorrect and separates the first two exons from the remaining exons of the OCA2 gene. We amplified a canine OCA2 cDNA fragment by RT-PCR and determined the correct full-length mRNA sequence (LT844587.1). Variants in the OCA2 gene cause oculocutaneous albinism type 2 (OCA2) in humans, pink-eyed dilution in mice, and similar phenotypes in corn snakes, medaka and Mexican cave tetra fish. We therefore conclude that the observed oculocutaneous albinism in German Spitz is most likely caused by the identified variant in the 5'-splice site of the first intron of the canine OCA2 gene.

Albinism, Oculocutaneous

Further observations on rangiferine brucellosis in Alaskan carnivores.

Antibodies against rangiferine brucellosis, Brucella suis type 4, are commonly found in the serum of various domestic and wild alaskian carnivores which feed on caribou, Rangifer tarandus granti, arctic Alaska. Sled dogs from five native villages on the range of the Artic caribou herd, but not from two villages on the the range of the Porcupine caribou herd, are commonly infected. Wolves (Canis lupus) and red foxes (Vulpes fulva) are less commonly infected. About 90% of the grizzly bears (Ursus arctos horribilis) associated with the Artic caribou herd and 30% of those associated with the porcupine caribou herd show serologic signs of exposure to Brucella, presumalby the enzootic strain present in Alaska caribou. This is the first evidence of natural Brucella infection in bears. It is concluded that infection of predators by enzootic strains of Brucella present in prey species (e.g., ruminants) is common to many areas of the world. Evidence from the literature and unpublished experimental data suggest that such infections may intefere with reproduction in wild species, but additional study is needed to clearly resolve this question.

Agglutination Tests

Successful protection of humans exposed to rabies infection. Postexposure treatment with the new human diploid cell rabies vaccine and antirabies serum.

Forty-five persons severely bitten by rabid dogs and wolves in Iran were treated after exposure with a new rabies vaccine produced in cultures of human diploid cells. All except one also received one injection of rabies immune serum. This treatment, in contrast to past experience with other vaccines, resulted in protection of all individuals against rabies. Thus, almost a century after the postexposure treatment of humans was initiated, an effective tool for protecting man against rabies has finally been developed.

Adolescent

Prevalence of Dirofilaria immitis in a stray dog population in western New York.

In a survey in 1973 undertaken to determine the prevalence of Dirofilaria immitis in 100 dogs from a public pound in western New York, 2 dogs were found to be positive. Microfilariae of Dipetalonema sp were not present. Hearts and pulmonary blood vessels were dissected for adult worms, and blood samples were examined for microfilariae, using a modified Knott technique. Although adult heartworms and microfilariae have been recovered from wild foxes, coyotes, and wolves in New York, this is the first report of heartworm in domesticated dogs in the state.

Animals