Atypical multiple myeloma in 3 young dogs.

Three dogs under 12 months old were diagnosed with atypical multiple myeloma (MM), having an aggressive multifocal anaplastic round cell sarcoma in bone marrow, viscera, and/or peripheral blood, which were confirmed by cytology and immunohistochemistry to be of plasma cell origin. The intramedullary sarcomas caused myelophthisis, osteolysis, and hypercalcemia. Complete or free light chain monoclonal gammopathy in the serum and/or urine was demonstrated by protein electrophoresis and immunofixation. The polymerase chain reaction for antigen receptor rearrangement assay performed on 2 cases identified a clonally rearranged immunoglobulin gene. Neoplastic cells lacked expression of CD45, CD3, CD18, CD21, CD34, and MHCII by flow cytometry. Immunohistochemistry revealed MUM1 immunoreactivity of the neoplastic cells. Combining all data, the diagnosis was MM. An aggressive form of MM in young dogs should be a differential diagnosis for patients with an immunoglobulin-productive, B cell-clonal, CD45-negative, MUM1-positive discrete cell neoplasm arising from the bone marrow.

Complete Blood Counts and Blood Smear Analyses in 312 Diabetic Dogs (2007-2017).

Diabetes mellitus is a common endocrinopathy in dogs that has been associated with various biochemical changes and comorbid diseases, but hematologic abnormalities have been rarely reported. The aim of this retrospective study was to evaluate complete blood count and blood smear alterations and to describe their relationship with, and incidence of comorbid diseases in, diabetic dogs. Three-hundred twelve diabetic dogs, 286 dogs diagnosed with systemic, nondiabetic illnesses, and 506 healthy dogs were identified during the study period. Groups were compared using contingency tables and logistic regression. Associations between statistically significant complete blood count and blood smear alterations and comorbidities were evaluated using multivariable analysis. High-grade codocytosis and anisocytosis were identified more frequently in diabetic dogs, whereas high-grade reactive lymphocytosis and keratocytosis were identified less frequently (P < .001). Diabetic dogs with high-grade codocytosis had lower red blood cell, hemoglobin, hematocrit and higher white blood cell counts (P < .001). Diabetic ketoacidosis was diagnosed more frequently in diabetic dogs with high-grade codocytosis when compared with those with low-grade codocytosis (P < .001) or when compared with any other cell morphologic alterations. This study suggests that blood smear analysis should be a routine part of the evaluation of diabetic dogs.

RNA-Seq based transcriptome of whole blood from immunocompetent pigs (Sus scrofa) experimentally infected with Mycoplasma suis strain Illinois.

Pigs are popular animal models in biomedical research. RNA-Seq is becoming the predominant tool to investigate transcriptional changes of the pig's response to infection. The high sensitivity of this tool requires a strict control of the study design beginning with the selection of healthy animals to provide accurate interpretation of research data. Pigs chronically infected with Mycoplasma suis often show no obvious clinical signs, however the infection may affect the validity of animal research. The goal of this study was to investigate whether or not this silent infection is also silent at the host transcriptional level. Therefore, immunocompetent pigs were experimentally infected with M. suis and transcriptional profiles of whole blood, generated by RNA-Seq, were analyzed and compared to non-infected animals. RNA-Seq showed 55 differentially expressed (DE) genes in the M. suis infected pigs. Down-regulation of genes related to innate immunity (tlr8, chemokines, chemokines receptors) and genes containing IFN gamma-activated sequence (gbp1, gbp2, il15, cxcl10, casp1, cd274) suggests a general suppression of the immune response in the infected animals. Sixteen (29.09%) of the DE genes were involved in two protein interaction networks: one involving chemokines, chemokine receptors and interleukin-15 and another involving the complement cascade. Genes related to vascular permeability, blood coagulation, and endothelium integrity were also DE in infected pigs. These findings suggest that M. suis subclinical infection causes significant alterations in blood mRNA levels, which could impact data interpretation of research using pigs. Screening of pigs for M. suis infection before initiating animal studies is strongly recommended.

Microscopy and genomic analysis of Mycoplasma parvum strain Indiana.

Mycoplasma parvum [Eperythrozoon parvum] is the second hemotrophic mycoplasma (hemoplasma) described in pigs. Unlike M. suis, its closest phylogenetic relative, M. parvum, is considered a non-pathogenic bacterium in this host species. Natural infection of a domestic, 6-month-old splenectomized pig with M. parvum strain Indiana is described herein. Light and scanning electron microscopy of the bacteria were performed in addition to whole genome sequencing, analysis, and comparison to the genome of M. suis strain Illinois. Neither clinical signs nor anemia were observed during the infection. Microscopy analyses revealed coccoid to rod- shaped organisms varying from 0.2 to 0.5 µm; they were observed individually or in short chains by both light and electron microscopy, however less than 30% of the red blood cells were infected at peak bacteremia. The single circular chromosome of M. parvum was only 564 395 bp, smaller than M. genitalium, previously considered the tiniest member of the Mollicutes. Its general genomic features were similar to others in this class and species circumscription was verified by phylogenomic analysis. A gene-by-gene comparison between M. suis and M. parvum revealed all protein coding sequences (CDS) with assigned functions were shared, including metabolic functions, transporters and putative virulence factors. However, the number of CDS in paralogous gene families was remarkably different with about half as many paralogs in M. parvum. The differences in paralogous genes may be implicated in the different pathogenic potential of these two species, however variable gene expression may also play a role. Both are areas of ongoing investigation.

Hemotropic Mycoplasmas (Hemoplasmas) in Free-Ranging Azara's Agoutis (Dasyprocta azarae) from an Urban Area of Southern Brazil.

Hemotropic mycoplasmas (hemoplasmas) are opportunistic bacteria that attach to the erythrocyte surface, causing infectious anemia in several mammalian species, including rodents. Studies surveying native Azara's agoutis (Dasyprocta azarae) in Brazil are lacking. Accordingly, the present study aimed to assess hemoplasmas infection in free-ranging agoutis from an urban environmental conservation area in Curitiba, southern Brazil. Overall, 11/35 (31.43%) agoutis were positive to hemoplasmas by quantitative PCR (cycle threshold≤34.4). Sequencing of the 16S ribosomal RNA gene indicated Mycoplasma haemomuris infection, closely related to M. haemomuris subsp. ratti, suggesting hemoplasma transmission from urban rats to agoutis. Because the main route of M. haemomuris transmission has been direct rodent-to-rodent infection, the relatively lower positivity that we detected may be the result of low intraspecies contact due to the smaller social units of agoutis, generally consisting of two to four individuals, and low interspecies contact due to only sporadic agouti-rat interactions in urban settings, compared with other rodent species interactions. Further studies should be conducted to determine whether the hemoplasma infection that we found can cause clinical onset and life-threatening anemia in agoutis.

Complete genome sequence of Mycoplasma haemocanis strain Illinois.

Mycoplasma haemocanis is a blood pathogen that may cause acute disease in immunosuppressed or splenectomized dogs. The genome of the strain Illinois is a single circular chromosome with 919,992 bp and a GC content of 35%. Analyses of the M. haemocanis genome will provide insights into its biology and in vitro cultivation requirements.

Genome sequence of "Candidatus Mycoplasma haemolamae" strain purdue, a red blood cell pathogen of alpacas (Vicugna pacos) and llamas (Lama glama).

We report the complete genome sequence of "Candidatus Mycoplasma haemolamae," an endemic red-cell pathogen of camelids. The single, circular chromosome has 756,845 bp, a 39.3% G+C content, and 925 coding sequences (CDSs). A great proportion (49.1%) of these CDSs are organized into paralogous gene families, which can now be further explored with regard to antigenic variation.

Complete genome sequence of Mycoplasma wenyonii strain Massachusetts.

Mycoplasma wenyonii is a hemotrophic mycoplasma that causes acute and chronic infections in cattle. Here, we announce the first complete genome sequence of this organism. The genome is a single circular chromosome with 650,228 bp and G+C% of 33.9. Analyses of M. wenyonii genome will provide insights into its biology.

Mycoplasma haemocanis--the canine hemoplasma and its feline counterpart in the genomic era.

Mycoplasma haemocanis is a hemotrophic mycoplasma (hemoplasma), blood pathogen that may cause acute disease in immunosuppressed or splenectomized dogs. The genome of the strain Illinois, isolated from blood of a naturally infected dog, has been entirely sequenced and annotated to gain a better understanding of the biology of M. haemocanis. Its single circular chromosome has 919 992 bp and a low G + C content (35%), representing a typical mycoplasmal genome. A gene-by-gene comparison against its feline counterpart, M. haemofelis, reveals a very similar composition and architecture with most of the genes having conserved synteny extending over their entire chromosomes and differing only by a small set of unique protein coding sequences. As in M. haemofelis, M. haemocanis metabolic pathways are reduced and apparently rely heavily on the nutrients afforded by its host environment. The presence of a major percentage of its genome dedicated to paralogous genes (63.7%) suggests that this bacterium might use antigenic variation as a mechanism to evade the host's immune system as also observed in M. haemofelis genome. Phylogenomic comparisons based on average nucleotide identity (ANI) and tetranucleotide signature suggest that these two pathogens are different species of mycoplasmas, with M. haemocanis infecting dogs and M. haemofelis infecting cats.

Hemophagocytic syndrome associated with large granular lymphoma in an adult dog.

Hemophagocytic syndrome (HPS) is a rare disorder characterized by dysregulation of the immune response resulting in uncontrolled activation of macrophages with exacerbated phagocytosis of host cells. In dogs, the criteria for diagnosis include the presence of pancytopenia or bicytopenia in the peripheral blood and >2% hemophagocytic macrophages in bone marrow aspirates. When HPS is associated with lymphoma, it is called lymphoma-associated hemophagocytic syndrome (LAHS). Here, we present a case of a 4 ½-year-old female spayed Old English Mastiff that presented with severe thrombocytopenia, mild anemia, mild to moderate leukopenia, and large granular lymphocytes (LGLs) in the peripheral blood. The patient had enlarged lymph nodes with many LGLs seen cytologically, leading to the interpretation of LGL lymphoma. Bone marrow displayed numerous LGLs that stained strongly for CD3 but did not show immunoreactivity to CD4 or CD8, and PCR for antigen receptor rearrangement analysis confirmed a clonal T-cell receptor gamma gene rearrangement. The presence of ~3.5% hemophagocytes present on the bone marrow evaluation raised concern for HPS and, more specifically, LAHS. HPS and LAHS are challenging to diagnose and require many criteria to be fulfilled before a definitive diagnosis can be made; the low number of cases in the literature makes this even more challenging in dogs. This case represents secondary LAHS due to LGL lymphoma in a dog.

Complete genome sequences of two hemotropic mycoplasmas, Mycoplasma haemofelis strain Ohio2 and Mycoplasma suis Strain Illinois.

We report the complete and fully assembled genomes of Mycoplasma haemofelis strain Ohio2 and Mycoplasma suis strain Illinois, which are the first available genomes of these uncultivatable hemoplasma species. The single circular chromosomes of 1,152,484 bp and 742,431 bp for M. haemofelis and M. suis, respectively, are typical of mycoplasma species, having reduced size and low G+C content (38.8% for M. haemofelis and 31.1% for M. suis). Their metabolic pathways are reduced, with evidence of adaption to the blood environment.

Genome of Mycoplasma haemofelis, unraveling its strategies for survival and persistence.

Mycoplasma haemofelis is a mycoplasmal pathogen (hemoplasma) that attaches to the host's erythrocytes. Distributed worldwide, it has a significant impact on the health of cats causing acute disease and, despite treatment, establishing chronic infection. It might also have a role as a zoonotic agent, especially in immunocompromised patients. Whole genome sequencing and analyses of M. haemofelis strain Ohio2 was undertaken as a step toward understanding its survival and persistence. Metabolic pathways are reduced, relying on the host to supply many of the nutrients and metabolites needed for survival. M. haemofelis must import glucose for ATP generation and ribose derivates for RNA/DNA synthesis. Hypoxanthine, adenine, guanine, uracil and CMP are scavenged from the environment to support purine and pyrimidine synthesis. In addition, nicotinamide, amino acids and any vitamins needed for growth, must be acquired from its environment. The core proteome of M. haemofelis contains an abundance of paralogous gene families, corresponding to 70.6% of all the CDSs. This "paralog pool" is a rich source of different antigenic epitopes that can be varied to elude the host's immune system and establish chronic infection. M. haemofelis also appears to be capable of phase variation, which is particularly relevant to the cyclic bacteremia and persistence, characteristics of the infection in the cat. The data generated herein should be of great use for understanding the mechanisms of M. haemofelis infection. Further, it will provide new insights into its pathogenicity and clues needed to formulate media to support the in vitro cultivation of M. haemofelis.

A case of acute monocytic leukemia (AMoL or AML-M5) in an adult FeLV/FIV-positive cat.

A 7-year-old castrated male domestic shorthair cat was presented for evaluation of decreased appetite and respiratory signs. A CBC run on presentation revealed severe nonregenerative anemia, thrombocytopenia, and leukocytosis characterized by a prominent population of blasts, having morphologic features suggestive of a monocytic lineage. The cat tested positive for FIV, FeLV, Mycoplasma haemominutum, and only mild abnormalities were identified on the chemistry panel. Bone marrow biopsies were obtained to investigate the bicytopenia and the possibility of a hematopoietic neoplasm. Although the bone marrow aspirate was nondiagnostic, the core biopsy was markedly hypercellular with a population of blasts, largely replacing the normal hematopoietic tissue. Immunohistochemical staining revealed that the blasts were CD3-negative, Pax5-negative, dimly CD18-positive, and moderately positive for Iba1. These findings, in addition to the prominent monocytic differentiation seen in peripheral blood, supported a diagnosis of acute monocytic leukemia. Palliative antiviral and antibiotic treatment and blood transfusion were performed. The patient was discharged on his fourth day of hospitalization. However, 15 days following discharge, the cat was euthanized due to the worsening of his systemic signs. This report discusses the classifications of myeloid leukemias, implications of infectious diseases in the pathogenesis of neoplasia in cats, and the use of Iba1, a "pan-monocytic/histiocytic" marker, in the diagnosis of acute leukemia.

Brucella canis endophthalmitis in 3 dogs: clinical features, diagnosis, and treatment.

OBJECTIVE: To describe historical, clinical and diagnostic features of dogs with Brucella canis endophthalmitis and the response to medical therapy. ANIMALS STUDIED: Three dogs with naturally acquired B. canis endophthalmitis. PROCEDURE: Dogs were treated symptomatically with topical ophthalmic anti-inflammatories and a novel antimicrobial protocol that included doxycycline, enrofloxacin, rifampin and streptomycin. RESULTS: All dogs presented with chronic or recurrent uveitis in the absence of overt systemic disease. Clinical ophthalmologic abnormalities were unilateral in each dog and included mild-to-moderate anterior uveitis, iris hyperpigmentation, marked vitreal infiltrates, and multifocal chorioretinitis. Dogs were diagnosed with canine brucellosis serologically and by blood culture (n = 2 dogs) or polymerase chain reaction of aqueous humor and blood (n = 1 dog). Active ocular inflammation resolved in all dogs during treatment, with preservation of vision in 2 dogs. Following treatment, B. canis could not be cultured from blood samples and serological values declined with seronegativity achieved in all dogs after a median of 96 weeks (range: 36-112 weeks) of therapy. CONCLUSIONS: Brucella canis infection should be included in the differential diagnosis for dogs with intraocular inflammation, regardless of previous history or neuter status. This is the first report of apparently successful medical therapy of canine brucellosis with ocular involvement.

Diagnosis and treatment of Babesia odocoilei in captive reindeer (Rangifer tarandus tarandus) and recognition of three novel host species.

Two captive reindeer (Rangifer tarandus tarandus) at a New York zoological institution were diagnosed with Babesia odocoilei. Clinical signs consistent with acute babesiosis included fever, hemoglobinuria, and hemolytic anemia. Both episodes were precipitated by stressful events that may have compromised their immunocompetence. The diagnosis was confirmed by visualization of intraerythrocytic parasites on stained blood smears, polymerase chain reaction, and speciation of the Babesia by sequencing a hypervariable region of the 18S rRNA gene. One reindeer died with gross and histopathologic lesions, including pigmentary nephrosis with severe acute tubular degeneration and necrosis secondary to intravascular hemolysis. A second reindeer was successfully treated with supportive care and an antiprotozoal, imidocarb dipropionate (Imizol, 12%, Schering-Plough Animal Health, Union, New Jersey 07083, USA) at 3 mg/kg s.c. or i.m. s.i.d. on days 1, 2, 6, 9, and 21. Two other reindeer in the exhibit tested negative for Babesia by polymerase chain reaction but were treated with imidocarb dipropionate as prophylaxis while final testing results were pending. Additionally, B. odocoilei was identified in three novel asymptomatic host species within the collection: yak (Bos grunniens), muntjac (Muntiacus reevesi), and markhor goat (Caprafalconeri). Due to the high morbidity and mortality associated with acute babesiosis, captive reindeer should receive tick prevention, be tested for subclinical infections in endemic areas, and receive aggressive treatment for acute infections when clinical babesiosis is suspected.

Survey of feline leukemia virus and feline coronaviruses in captive neotropical wild felids from Southern Brazil.

A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center (Refúgio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44 animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas reverse transcriptase-PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these species in Southern Brazil.

Chronic myelogenous leukemia in a great horned owl (Bubo virginianus).

A free-ranging adult female great horned owl (Bubo virginianus) was presented to the Wildlife Medical Clinic at the University of Illinois after being observed with anorexia and decreased activity. A severe leukocytosis (212 400 cells/microl), primarily comprised of mature heterophils, was found at presentation. Results of various diagnostic tests including radiographs, Chlamydophila serologic testing, measurement of Aspergillus antibody and antigen titers, plasma protein electrophoresis, fecal culture and acid-fast staining, coelioscopy, endoscopy, tracheoscopy, exploratory coelomotomy, nuclear scintigraphy, tissue cultures, bone marrow biopsy, and histopathology revealed no underlying cause for the persistent leukocytosis. No response to treatment with antibiotics or antifungal agents was observed, although a transient, significant decrease in the leukocyte count (6200 cells/microl) was observed after treatment with fenbendazole. A presumptive diagnosis of chronic myelogenous leukemia was made based on 3 factors: disease duration of greater than 3 months, a lack of identifiable foci of inflammation, and a lack of response to conventional therapy. The diagnosis was confirmed based on postmortem examination and testing 177 days after initial presentation.

Genomic profile of Brazilian methicillin-resistant Staphylococcus aureus resembles clones dispersed worldwide.

PURPOSE: Comparative genomic analysis of strains may help us to better understand the wide diversity of their genetic profiles. The aim of this study was to analyse the genomic features of the resistome and virulome of Brazilian first methicillin-resistant Staphylococcus aureus (MRSA) isolates and their relationship to other Brazilian and international MRSA strains. METHODOLOGY: The whole genomes of three MRSA strains previously isolated in Vitória da Conquista were sequenced, assembled, annotated and compared with other MRSA genomes. A phylogenetic tree was constructed and the pan-genome and accessory and core genomes were constructed. The resistomes and virulomes of all strains were identified.Results/Key findings. Phylogenetic analysis of all 49 strains indicated different clones showing high similarity. The pan-genome of the analysed strains consisted of 4484 genes, with 31 % comprising the gene portion of the core genome, 47 % comprising the accessory genome and 22 % being singletons. Most strains showed at least one gene related to virulence factors associated with immune system evasion, followed by enterotoxins. The strains showed multiresistance, with the most recurrent genes conferring resistance to beta-lactams, fluoroquinolones, aminoglycosides and macrolides. CONCLUSIONS: Our comparative genomic analysis showed that there is no pattern of virulence gene distribution among the clones analysed in the different regions. The Brazilian strains showed similarity with clones from several continents.

Hemoplasma infection in HIV-positive patient, Brazil.

Hemotrophic mycoplasmas infect a variety of mammals. Although infection in humans is rarely reported, an association with an immunocompromised state has been suggested. We report a case of a Mycoplasma haemofelis-like infection in an HIV-positive patient co-infected with Bartonella henselae.

Prevalence and risk factors for hemoplasmas in domestic cats naturally infected with feline immunodeficiency virus and/or feline leukemia virus in Rio de Janeiro--Brazil.

The aim of this study was to determine the prevalence and risk factors for Mycoplasma haemofelis (Mhf) and 'Candidatus Mycoplasma haemominutum' (Mhm) infections in domestic cats tested for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Based on serological testing, cats were grouped as i) FIV-positive (n=25); ii) FeLV-positive (n=39); iii) FIV/FeLV-positive (n=8); and iv) FIV/FeLV-negative (n=77). Complete blood counts were followed by DNA extraction, species-specific polymerase chain reaction (16S rRNA gene) for Mhf and Mhm and Southern blotting for all animals. Mhf DNA was found in 4.0, 2.6, 12.5 and 7.8% of the cats from groups i, ii, iii and iv, respectively, while 32, 5.1, 50 and 5.2% of these animals had an Mhm infection. Cats with FIV (OR=4.25, P=0.009) and both FIV and FeLV (OR=7.56, P=0.014) were at greater risk of being hemoplasma infected than retroviral-negative cats, mainly due to Mhm infection (OR=8.59, P=0.001 and OR=18.25, P=0.001, respectively). Among pure-breed cats, FIV-positive status was associated with hemoplasma infection (OR 45.0, P=0.001).