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This study aimed to determine the effect of rice husk fluidization and variation in the equivalence ratio of bubbling fluidized bed gasifiers without sand bed materials. It also aimed to improve the fluidization quality by reducing the diameter of rice husks. Therefore, the bulk density increases, whereas voidage decreases, both of which are the main parameters for improving the quality of fluidization in solid particles. Experiments were carried out at a velocity of 0.82 m/s, by varying the equivalent ratios ranging from 0.20 to 0.35, and analyzing the syngas composition, cold gas and carbon conversion efficiencies, lower heating value, and temperature distribution. An equivalence ratio of 0.30 was obtained for a bubbling fluidized gasifier with syngas compositions of 7.415%, 15.674%, 3.071%, 17.839%, and 56.031% for H2, CO, CH4, CO2, and N2, respectively. Under these conditions, we obtained cold gas and carbon conversion efficiencies and a lower heating value of 31.340%, 37.120%, and 3.881 MJ/Nm3, respectively.

At this time the research related to the effect of magnetostriction for underwater applications is still limited. Acoustic or sound waves are more easily propagated underwater than electromagnetic waves or light. An acoustic wave or sound can be generated by utilizing the magnetostriction effect, where this effect occurs when a rod of ferromagnetic material such as iron or nickel is magnetized and interacts with another magnetic field, resulting in vibration of the metal rod. This research aims to design and realize a magnetostriction generator as an acoustic wave generator at a frequency of 1 to 10 kHz for underwater applications, consisting of: a tuned LC oscillator circuit, and a ferromagnetic metal rod which is magnetized using a dc voltage. The results of measurements and testing of the magnetostriction generator show: (1) if it is equipped with a membrane, can work to emit an acoustic wave or sound at a frequency of ± 8.62 kHz in the air up to a distance of 15 cm without distortion with an average amplitude decrease of ± 0.648 dB for each the distance increased by 1 cm, and (2) if equipped with a membrane and enclosed in a waterproof casing, capable of transmitting the acoustic waves at a frequency of ± 8.31 kHz underwater up to a distance of 7 cm without distortion with an average amplitude decrease of ± 4.217 dB for each the distance up 1 cm. Overall the magnetostriction generator designed can work to generate and transmit the acoustic waves or sound underwater, as expected.

To find out the location of an aircraft accident from an ELT signal beam of 121.5 MHz, search and rescue (SAR) officers or civil aviation authorities need an ELT signal receiver device that works at the same frequency, which is 121.5 MHz. The purpose of this research is to produce an ELT signal receiver with direct receiver method at a frequency of 121.5 MHz which is simpler, requires fewer RF devices, and is according to specifications for civil aviation. In this research, the design and realization of ELT signal receivers are carried out using the direct receiver method, including: a BPF and a RF amplifier as a direct receiver circuit, an AM demodulator, and an audio amplifier. Based on the results of measurements and tests that have been done, the direct receiver circuit can receive an ELT signal at a frequency of 121.5 MHz in the range of 121.330 to 121.617 MHz with -40.35 dB sensitivity, the AM demodulator can demodulate the audio signal at a frequency of 346.6 Hz, and the audio signal at a frequency of 337.6 Hz as a pointer to an ELT signal can be heard through the loudspeaker. The results of this study are ELT signal receivers with direct receiver method at a frequency of 121.5 MHz which are according to specifications for civil aviation.

A Radio detection and ranging (radar) is used to detect and track objects in the air and on land, one type of the radar that is known is a passive radar. The radio direction finder (RDF) as an element of a passive radar is used to determine the direction angle, based on a radio waves that emitted by an object. The purpose of this study is to produce a RDF that works on HF bands as passive radar elements. In this study a RDF has been produced, which consists: a receive antenna in the form of a dipole λ/8 and a yagi 5 elements with horizontal polarization, a tuning circuit, an AM signal detector, an audio amplifier, and equipped with a level signal display in the form of a S meter and a loudspeaker. The test results for outside and inside the room at a certain time, the RDF can detect the angle of direction of horizontal plane of an object or AM transmitter from 0° to 360° in the frequency range between 4.02 to 10.76 MHz. The results of detecting the angle of direction of an object or a transmitting radio waves in the HF band, in the form of a signal level are then plotted on the polar diagram of the horizontal plane, and the sound can be heard through the loudspeaker. Furthermore, the results of this study can be used for the development of a RDF independently, and its implementation in the passive radar.

An ELT is a beacon indicating the location of distress or crash of aircraft on land or sea. The ELT is placed in an airplane, which is a transmitter with low transmit power and an antenna. The ELT transmitter has been widely used for flights at the frequency of 121.5 MHz (civilian) and 243 MHz (military). The purpose of this study is to produce ELT signal generators at the 121.5 MHz frequency, with low-cost devices that meet specifications for civil aviation. In this research has been produced a design of an ELT signal generator at the frequency of 121.5 MHz, consist of a crystal oscillator, a sawtooth generator, a VCO, and an amplitude modulator. Based on the measurement and test results, the sawtooth generator generated a sawtooth signal at the frequency of 1.8 to 3.6 Hz, the VCO generated a sinusoidal signal at the frequency of 316 to 366 Hz, the RF generator generated a sinusoidal at the frequency of 121.505 MHz. The ELT signal generator was able to generate AM signals at the minimum amplitude deviation of 100 mVpp to maximum one of 156 mVpp, the modulation index (m) of 21.875%, and the frequency of 121.505 MHz. The results of this study are ELT signal generators at the frequencies of 121.5 MHz that are in accordance with specifications for civil aviation.

Black box yang terdiri cockpit voice recorder dan flight data recorder dapat dideteksi lokasinya karena dilengkapi dengan beacon black box locator acoustic 37,5 kHz atau pinger yang memancarkan sinyal akustik 37,5 kHz. Untuk mendeteksi sinyal pinger yang mempunyai frekuensi 37,5 kHz, digunakan detektor sinyal pinger, yang mampu mendeteksi gelombang ultrasonic di dalam air pada frekuensi 37,5 1 kHz. Makalah ini membahas perancangan pembangkit dan detektor sinyal pinger 37,5 kHz. Rancangan pembangkit sinyal pinger untuk membangkitkan sinyal pinger dengan frekuensi keluaran 37,5 1 kHz, terdiri rangkaian osilator, pulse generator, pulse inverter, modulator OOK, dan audio amplifier. Rancangan detektor sinyal pinger untuk mendeteksi sinyal pinger dengan frekuensi 37,5 1 kHz, terdiri rangkaian band pass filter, demodulator OOK, audio amplifier, dan loudspeaker. Berdasarkan hasil pengujian, pembangkit sinyal pinger mampu membangkitkan sinyal pinger dengan amplitudo 12,2 Vp-p, frekuensi 37,69 kHz, lebar pulsa (kondisi on sebesar 9,8 ms), dan PRF sebesar 1 s sedangkan detektor sinyal pinger mampu mendeteksi sinyal pinger 37,5 kHz dan menghasilkan keluaran berupa pulsa dengan amplitudo 9 Vp-p, lebar pulsa 14 ms, dan PRF sebesar 1 s, serta suara yang dihasilkan terdengar jelas setiap interval 1 s.

ABSTRAK Dalam pencarian suatu obyek di bawah air dibutuhkan metode khusus yang digunakan dengan suara (gelombang akustik), dikarenakan suara dapat merambat dalam jarak jauh pada air. Frekuensi yang dapat diterima oleh SONAR bergantung pada alat penerima yang mengubah suara menjadi sinyal elektrik agar dapat mendeteksi frekuensi, alat penerima ini dinamakan hydrophone. Dalam penelitian ini dibuat perancangan prototipe hydrophone, yang tersusun dari transducer (akustik-elektrik) dan penguat. Berdasarkan perancangan yang telah dibuat prototipe hydrophone mampu menerima (menangkap) gelombang akustik pada range frekuensi 100 Hz – 60 kHz, dengan menggunakan transducer berupa condenser microphone yang diselimuti oleh bahan karet tipis. Prototipe hydrophone tanpa pelindung anti air, dengan pelindung anti air, dan di dalam air dapat menerima (menangkap) gelombang akustik. Kata Kunci: SONAR, hydrophone, transducer ABSTRACT In finding an object beneath the water there has to be a specific method used sound (accoustic wave) in keeping with it can apread in long distance in the depth of water. The frequency which can be detected by sonar system, counts on the receiver which converts sound to electric signal so that the frequency can be detected and it is called hydrophone. In this research, there is hydrophone prototype which piled up by tranducer and amplifier. Based on the trial, hydrophone prototype can detect accoustic wave in range 100 Hz-60 kHz with the condensor microphone as a transducer covered of thin rubber material. Besides, hydrophone prototype with or without water resistor is still capable to detect acoustic wave. Keywords: SONAR, hydrophone, transducer

ABSTRAK Pingers transmitter berfungsi untuk memancarkan sinyal atau getaran pulsa akustik pada black box. Frekuensi sinyal yang dipancarkan sebesar 37,5 kHz yang dimodulasikan oleh pulsa dengan durasi 10 ms setiap interval 1 second. Modulasi yang digunakan adalah modulasi on off keying. Dalam penelitian ini dibuat perancangan pingers transmitter yang tersusun atas rangkaian osilator, timer, inverter, switch dan rangkaian amplifier. Frekuensi 37,5 kHz tersebut dibangkitkan oleh rangkaian osilator colpitts, sedangkan lebar pulsa dengan durasi 10 ms dan pengulangan pulsa setiap interval 1 second dibangkitkan oleh rangkaian timer. Berdasarkan perancangan yang telah dibuat, output sinyal yang dihasilkan oleh prototipe pingers transmitter tersebut sebesar 37,69 kHz dengan lebar pulsa 9,8 ms setiap interval 1 second. Kata kunci: Black box, pingers transmitter, on off keying. ABSTRACT Pingers transmitter is used to emits a signal or pulse of acoustic vibrations in black box. The frequency of the transmitted signal is 37.5 kHz which is modulated by pulses with a duration of 10 ms every interval 1 second. The modulation that used this research is on off keying modulation. In this research, made the design of pingers transmitter which arrange of the oscillator circuit,timer, inverter, switch, and amplifier circuit. The 37.5 kHz frequency is generated by the colpitts oscillator circuit, while the pulse width with a duration of 10 ms,and each pulse repetition interval of 1 second generated by timer circuit. Based on the design that have made, signal output from the pingers transmitter prototype is 37.69 kHz, with the pulse width 9.8 ms every 1 second interval. Keywords: Black box, pingers transmitter, on off keying.

ABSTRAK Keandalan tenaga listrik didefinisikan sebagai peluang dari suatu peralatan untuk beroperasi sesuai dengan fungsinya dalam suatu selang waktu tertentu dan dalam suatu kondisi operasi tertentu, sehingga dapat memenuhi kebutuhan listrik konsumen. Keandalan ketersediaan daya suatu sistem pembangkit dapat diketahui berdasarkan indeks keandalan yaitu LOLP (Loss of Load Probability) dan UE (Unserved Energy). Penelitian dilakukan dengan merancang 2 model konfigurasi sistem pembangkit berdasarkan nilai beban puncak, kemudian mengambil data laju kegagalan dan laju perbaikan dari masing-masing komponen sistem pembangkit yang selanjutnya dipakai untuk menghitung nilai FOR (Forced Outage Rate). Nilai FOR yang telah diperoleh digunakan untuk menghitung nilai indeks keandalan menggunakan metode segmentasi. Konfigurasi sistem pembangkit skenario ke-2 memiliki nilai indeks keandalan lebih kecil dibandingkan dengan konfigurasi sistem pembangkit skenario ke-1 yaitu dengan LOLP sebesar 0,0000088248 hari/tahun dan UE sebesar 0,0584 KWH/tahun dengan total perkiraan biaya pada konfigurasi ini sebesar Rp 587.567.108,00. Kata kunci: Keandalan, metode segmentasi, LOLP, FOR, UE ABSTRACT The reliability of electric power is defined as the chance of an apparatus to operate in accordance with its function in a certain interval of time and in a certain operating conditions, so as to meet the electricity needs of consumers. The reliability of a power generation system availability can be determined based on the reliability index is LOLP (Loss of Load Probability) and UE (Unserved Energy). The study was conducted by designing two models generating system configuration based on the value of the peak load, and then retrieve the data failure rate and the rate of repair of the individual components of the next generation system which is used to calculate the value FOR (Forced Outage Rate). FOR Value that has been obtained is used to calculate the reliability index using segmentation method. Generating system configuration scenarios to-2 has a reliability index value is smaller than the generating system configuration scenarios to-1 that is by LOLP of 0.0000088248 days / year and the EU amounted to 0.0584 KWH / year with a total estimated cost in this configuration Rp 587,567,108.00. Keywords: Reliability, segmentation method, LOLP, FOR, UE

Parelaphostrongylus tenuis causes ungulate morbidity and mortality in eastern and central North America, but no reference genome sequence exists to facilitate research. Here, we present a P. tenuis genome assembly and annotation, generated with PacBio and Illumina technologies. The assembly is 491 Mbp, with 7285 scaffolds and 185 kb N50., (© 2024 Tyler J. Garwood et al., published by Sciendo.)

Staphylococcus schleiferi and Staphylococcus coagulans , closely related bacterial species within the Staphylococcus genus, present a challenge in classification and diagnosis due to their close genetic proximity and overlapping phenotypic features. Moreover, our understanding of the virulence mechanisms in staphylococcal species, beyond the extensively studied Staphylococcus aureus , remains limited, underscoring the importance of using comparative data to enhance our insights into virulence within these bacterial species. This study employed a comprehensive approach, utilizing comparative genomics, to identify genomic distinctions between S. schleiferi and S. coagulans , aiming to address the challenges in the accurate classification and diagnosis of these organisms and identify unique features. Whole genome sequencing was performed on six clinical isolates, and their genomes were compared to identify variations in gene content and virulence factors. De novo assembly and annotation revealed two samples as S. coagulans and four samples as S. schleiferi . Analysis of the core genomes revealed conserved regions crucial for defining species identity, while accessory genomic elements contained unique genes, possibly impacting the pathogenicity of the species.

Staphylococcus schleiferi and Staphylococcus coagulans are opportunistic pathogens of animals and humans. They were previously classified as Staphylococcus schleiferi subs. schleiferi and Staphylococcus schleiferi subs. coagulans, respectively, and recently reclassified as separate species. S. coagulans, is frequently associated with dogs, whereas S. schleiferi is more commonly isolated from humans. Coagulase activity status is a defining characteristic of the otherwise closely related species. However, the use of coagulase tests originally developed to distinguish S. aureus from non-coagulase-producing staphylococci, for this purpose is questionable and the basis for their host preference has not been elucidated. In the current study, a putative coa gene was identified and correlated with coagulase activity measured using a chromogenic assay with human and bovine prothrombin (closely related to canine prothrombin). The results of the tests performed with human prothrombin showed greater reactivity of S. coagulans isolates from humans than isolates obtained from dogs with the same substrate. Our data suggest that unlike S. coagulans isolates from humans, isolates from dogs have more coagulase activity with bovine prothrombin (similar to canine prothrombin) than human prothrombin. Differences in nuc and 16s rRNA genes suggest a divergence in S. coagulans and S. schleiferi. Phenotypic and genotypic variation based on the number of IgG binding domains, and the numbers of tandem repeats in C-terminal fibronectin binding motifs was also found in protein A, and fibronectin-binding protein B respectively. This study identified a coa gene and associated phenotypic activity that differentiates S. coagulans and S. schleiferi and identified key phylogenetic and phenotypic differences between the species., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Sewid, Kania. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Staphylococcus pseudintermedius is an important cause of clinical infections in small-animal-veterinary medicine. Evolutionary changes of strains using multilocus sequence typing (MLST) have been observed among S. pseudintermedius in European countries and the United States. However, there are limited or no studies on the detection of methicillin resistant Staphylococcus pseudintermedius (MRSP) and predominating MLST strains in South Africa. Therefore, this study aimed to determine the molecular epidemiology of S. pseudintermedius in South Africa. Twenty-six, non-duplicate, clinical isolates from dogs were obtained as convenience samples from four provinces in South Africa. The Kirby Bauer disk diffusion test was used to determine antimicrobial susceptibility. We used Resfinder and the Comprehensive Antibiotic Resistance Database (CARD) to detect antimicrobial resistance genes. Virulence genes were identified using the virulence factor database and Basic Local Alignment Search Tool (BLASTN) on Geneious prime. geoBURST analysis was used to study relationships between MLST. Finally, the maximum likelihood phylogeny was determined using Randomized Axelerated Maximum Likelihood (RAxML). Twenty-three isolates were confirmed as S. pseudintermedius of which 14 were MRSP. In addition to β-lactam antimicrobials, MRSP isolates were resistant to tetracycline (85.7%), doxycycline (92.8%), kanamycin (92.8%), and gentamicin (85.7%). The isolates harbored antimicrobial resistance genes (tetM, ermB, drfG, cat, aac(6')-Ie-aph(2")-Ia, ant(6)-Ia, and aph(3')-III) and virulence genes (AdsA, geh, icaA, and lip). MLST analysis showed that ST2228, ST2229, ST2230, ST2231, ST2232, ST2318, ST2326 and ST2327 are unique sequence types in South Africa. Whereas, previously reported major STs including ST45, ST71, ST181, ST551 and ST496 were also detected. The geoBURST and phylogenetic analysis suggests that the isolates in South Africa are likely genetically related to isolates identified in other countries. Highly resistant MRSP strains (ST496, ST71, and ST45) were reported that could present challenges in the treatment of canine infections in South Africa. Hence, we have gained a better understanding of the epidemiology of MRSP in the African continent, the genes involved in resistance and virulence factors associated with these organisms., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Phophi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

We report here the genome sequences of 11 canine Staphylococcus pseudintermedius isolates from New York, New Hampshire, California, Pennsylvania, and Kansas. The sequencing information will enable spatial phylogenetic comparisons of staphylococcal species and other related species and will help in better understanding their virulence potential.

Foods that contain antibiotic residues have potential adverse health effects on consumers and provide selective pressure for the threat of antimicrobial resistance (AMR). This study's objective was to measure tetracycline, sulfonamide, and erythromycin residues in beef, eggs, and honey sold as "antibiotic-free" at farmers' markets in East Tennessee (East TN) in the United States (U.S.). Between July and September 2020, 36 "antibiotic-free" food products (9 beef, 18 egg, and 9 honey products) were purchased from East TN farmers' markets and tested for tetracycline, sulfonamide, and erythromycin residues using competitive enzyme-linked immunosorbent assays (cELISA). All beef, egg, and honey products had tetracycline residue; the median concentrations were 51.75, 30.25, and 77.86 µg/kg, respectively. Sulfonamide residue was present in every sample of beef. Of 18 eggs, 11 eggs had detectable sulfonamide residue; the median concentrations were 3.50 and 1.22 µg/kg in beef and eggs, respectively. Each sample of beef and honey contained erythromycin residue; the median concentrations were 3.67 and 0.68 µg/kg, respectively. Overall, the median concentrations of tetracycline, sulfonamide, and erythromycin residues were below the maximum residue levels (MRLs) set in the U.S. for beef and eggs. Thus, the beef and eggs sold as "antibiotic-free" in East TN farmers' markets can be considered safe for consumption. Safety determination for honey could not be made because MRLs have not been set for honey in the U.S. Because these residues should not be expected in "antibiotic-free" food products, it is important to further investigate the potential sources of these residues in these products.

Background: Understanding drivers of multidrug resistance (MDR) and methicillin resistance, which have increased among canine staphylococcal isolates, is essential for guiding antimicrobial use practices. Therefore, the objective of this study was to identify predictors of MDR and methicillin resistance among Staphylococcus spp. commonly isolated from canine clinical specimens., Methods: This retrospective study used records of canine specimens submitted to the University of Tennessee College of Veterinary Medicine Clinical Bacteriology Laboratory for bacterial culture and antimicrobial susceptibility testing between 2006 and 2017. Records from 7,805 specimens positive for the following Staphylococcus species were included for analysis: Staphylococcus pseudintermedius, Staphylococcus aureus, Staphylococcus coagulans (formerly Staphylococcus schleiferi subspecies coagulans ), and Staphylococcus schleiferi (formerly S. schleiferi subsp. schleiferi ). Generalized linear regression models were fit using generalized estimating equations (GEE) to identify predictors of MDR (defined as resistance to three or more antimicrobial classes) and methicillin resistance among these isolates., Results: Multidrug resistance (42.1%) and methicillin resistance (31.8%) were relatively common. Isolates from skeletal (joint and bone) specimens had the highest levels of MDR (51.3%) and methicillin resistance (43.6%), followed by cutaneous specimens (45.8% multidrug-resistant, 37.1% methicillin resistant). Staphylococcus species, specimen site, and clinical setting were significant ( p  < 0.01) predictors of both outcomes. Compared to S. pseudintermedius, S. schleiferi had higher odds of methicillin resistance, while S. coagulans and S. schleiferi had lower odds of MDR. The odds of both MDR and methicillin resistance for isolates from hospital patient specimens were significantly higher than those from referral patients for urine/bladder and otic specimens. Odds of MDR among isolates from skeletal specimens of hospital patients were also higher than those of referral patients., Conclusions: Staphylococcus isolates in this study had substantial levels of MDR and methicillin resistance. Differences in the odds of these outcomes between referral and hospital patient isolates did not persist for all specimen sites, which may reflect differences in diagnostic testing and antimicrobial use practices with respect to body site or system. Judicious antimicrobial use, informed by culture and susceptibility testing, is important to limit treatment failures and curb selection pressure., Competing Interests: Agricola Odoi is an Academic Editor for PeerJ., (©2023 Lord et al.)

In late 2019, a novel coronavirus began circulating within humans in central China. It was designated SARS-CoV-2 because of its genetic similarities to the 2003 SARS coronavirus (SARS-CoV). Now that SARS-CoV-2 has spread worldwide, there is a risk of it establishing new animal reservoirs and recombination with native circulating coronaviruses. To screen local animal populations in the United States for exposure to SARS-like coronaviruses, we developed a serological assay using the receptor binding domain (RBD) from SARS-CoV-2. SARS-CoV-2's RBD is antigenically distinct from common human and animal coronaviruses, allowing us to identify animals previously infected with SARS-CoV or SARS-CoV-2. Using an indirect enzyme-linked immunosorbent assay (ELISA) for SARS-CoV-2's RBD, we screened serum from wild and domestic animals for the presence of antibodies against SARS-CoV-2's RBD. Surprisingly prepandemic feline serum samples submitted to the University of Tennessee Veterinary Hospital were ∼50% positive for anti-SARS RBD antibodies. Some of these samples were serologically negative for feline coronavirus (FCoV), raising the question of the etiological agent generating anti-SARS-CoV-2 RBD cross-reactivity. We also identified several white-tailed deer from South Carolina with anti-SARS-CoV-2 antibodies. These results are intriguing, as cross-reactive antibodies toward SARS-CoV-2 RBD have not been reported to date. The etiological agent responsible for seropositivity was not readily apparent, but finding seropositive cats prior to the current SARS-CoV-2 pandemic highlights our lack of information about circulating coronaviruses in other species. IMPORTANCE We report cross-reactive antibodies from prepandemic cats and postpandemic South Carolina white-tailed deer that are specific for that SARS-CoV RBD. There are several potential explanations for this cross-reactivity, each with important implications to coronavirus disease surveillance. Perhaps the most intriguing possibility is the existence and transmission of an etiological agent (such as another coronavirus) with similarity to SARS-CoV-2's RBD region. However, we lack conclusive evidence of prepandemic transmission of a SARS-like virus. Our findings provide impetus for the adoption of a One Health Initiative focusing on infectious disease surveillance of multiple animal species to predict the next zoonotic transmission to humans and future pandemics.

Background: Mycobacteria are found in many environmental conditions and infect a variety of species, including rodents and rabbits. Guinea pigs are used experimentally as a model for Mycobacterium tuberculosis, but natural mycobacteriosis in guinea pigs has not been reported., Case Presentation: A 1.5-year-old female guinea pig was found acutely deceased with no premonitory illness. On gross post-mortem examination, multifocal to coalescing, raised, firm, pale tan nodules with discrete, irregular margins were noted over the surfaces of all lung lobes. Histopathology revealed nodules composed of clustered foamy macrophages and multinucleated giant cells containing numerous bacterial rods. Similar bacteria-laden macrophages were noted within sections of the liver, heart, palpebral conjunctiva, duodenum, and cecum. Polymerase chain reaction was performed on tissues collected during post-mortem examination. The 16S rRNA gene product was sequenced and was identical to the Mycobacterium genavense type strain., Conclusions: To the best of the author's knowledge, this report details the first documented case of Mycobacterium genvaense infection in a guinea pig and a follow up investigation of close-contact animals. Given their experimental susceptibility and this clinical case report, mycobacteriosis should be considered as a differential in guinea pigs exhibiting weight loss in the absence of other clinical signs. With the potential for zoonotic transmission in immunosuppressed individuals, precautions should be taken to safeguard human health in cases of guinea pigs with suspected M. genavense infection., (© 2022. The Author(s).)

Background: Multidrug- and methicillin-resistant staphylococci are both veterinary and public health concerns due to their zoonotic potential. Therefore, the objective of this study was to investigate patterns of antimicrobial, multidrug, and methicillin resistance among four Staphylococcus spp. commonly isolated from canine clinical specimens submitted to the Clinical Bacteriology Laboratory at the University of Tennessee College of Veterinary Medicine (UTCVM)., Methods: Results of antimicrobial susceptibility testing and mecA polymerase chain reaction (PCR) for isolates of four common Staphylococcus spp. isolates were obtained from the Bacteriology Laboratory at the UTCVM between 01/01/2006 and 12/31/2017. Cochran-Armitage trend test was used to assess temporal trends of antimicrobial resistance (AMR), multidrug resistance (MDR), and methicillin resistance. Kappa test of agreement was used to assess agreement between the results of PCR and disk diffusion tests., Results: Most of the 7805 isolates were S. pseudintermedius (6453 isolates), followed by S. coagulans (860), S. aureus (330), and S. schleiferi (162). Among S. pseudintermedius isolates, 45.5% were MDR, and 30.8% were methicillin-resistant (MRSP). There was a significant temporal increase in MRSP (p = 0.017). Chloramphenicol resistance increased among both MRSP and methicillin-susceptible (MSSP) isolates (p <  0.0001). Among S. aureus isolates, 40.9% were MDR, 37.4% were methicillin-resistant (MRSA), and the proportion of MRSA isolates increased significantly (p = 0.0480) over time. There was an increasing temporal trend in the proportion of MDR isolates among MSSP (p = 0.0022), but a decrease among MRSP (p <  0.0001) and MRSA (p = 0.0298). S. schleiferi had the highest percentage (56.9%) of methicillin-resistant isolates. Oxacillin disk diffusion was superior to cefoxitin for the detection of mecA-mediated resistance and had almost perfect agreement with mecA PCR assay for S. pseudintermedius (95.4% agreement, kappa (κ) = 0.904; p <  0.0001), S. coagulans (95.6%, κ = 0.913; p <  0.0001) and S. schleiferi (97.7%, κ = 0.945; p <  0.0001). However, cefoxitin disk diffusion was superior to oxacillin disk diffusion and had almost perfect agreement with mecA PCR assay for S. aureus (95.3%, κ = 0.834; p <  0.0001)., Conclusions: The levels of resistance and increasing temporal trends are concerning. These findings have implications for treatment decisions and public health due to the zoonotic potential of staphylococci. Continued surveillance and use of antibiograms to guide clinical decisions will be critical., (© 2022. The Author(s).)

Staphylococci have evolved numerous strategies to evade their hosts' immune systems. Some staphylococcal toxins target essential components of host innate immunity, one of the two main branches of the immune system. Analysis of the Staphylococcus pseudintermedius secretome using liquid chromatography mass spectrometry guided by genomic data, was used to identify an S. pseudintermedius exotoxin provisionally named SpEX. This exoprotein has low overall amino acid identity with the Staphylococcus aureus group of proteins named staphylococcal superantigen like proteins (SSLs) and staphylococcal enterotoxin- like toxin X (SEIX), but predictive modeling showed that it shares similar folds and domain architecture to these important virulence factors. In this study, we found SpEX binds to complement component C5, prevents complement mediated lysis of sensitized bovine red blood cells, kills polymorphonuclear leukocytes and monocytes and inhibits neutrophil migration at sub-lethal concentrations. A mutant version of SpEX, produced through amino acid substitution at selected positions, had diminished cytotoxicity. Anti-SpEX produced in dogs reduced the inhibitory effect of native SpEX on canine neutrophil migration and protected immune cells from the toxic effects of the native recombinant protein. These results suggest that SpEX likely plays an important role in S. pseudintermedius virulence and that attenuated SpEX may be an important candidate for inclusion in a vaccine against S. pseudintermedius infections., Competing Interests: I declare that we have a patent application, PCT/US2018/04681, entitled “Staphylococcus pseudointermedius virulence factor compositions” that includes the subject of this manuscript. The inventors are Stephen Kania, David Bemis and Mohamed Abouelkhair and the invention is owned by the University of Tennessee Research Foundation. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

The success of staphylococci as pathogens has been attributed, in part, to their ability to evade their hosts' immune systems. Although the proteins involved in evasion have been extensively studied in staphylococci affecting humans little characterization has been done with Staphylococcus pseudintermedius, an important cause of pyoderma in dogs. Staphylococcus aureus binder of immunoglobulin (Sbi) interferes with innate immune recognition by interacting with multiple host proteins. In this study, a S. pseudintermedius gene that shares 38% similarity to S. aureus Sbi was cloned from S. pseudintermedius strains representative of major clonal lineages bearing two paralogs of the protein. Binding of immunoglobulins and Fab and Fc fragments as well as interaction with complement was measured. S. pseudintermedius Sbi protein bound IgG from multiple species and canine complement C3, neutralized complement activity and bound to canine IgM and B cells. Evidence from this work suggests Sbi may play an important role in S. pseudintermedius immune evasion., Competing Interests: The authors have declared that no competing interests exist.

Light chain-associated amyloidosis is characterized by the extracellular deposition of amyloid fibrils in abdominothoracic organs, skin, soft tissue, and peripheral nerves. Phagocytic cells of the innate immune system appear to be ineffective at clearing the material; however, human light chain amyloid extract, injected subcutaneously into mice, is rapidly cleared in a process that requires neutrophil activity. To better elucidate the phagocytosis of light chain fibrils, a potential method of cell-mediated dissolution, amyloid-like fibrils were labeled with the pH-sensitive dye pHrodo red and a near infrared fluorophore. After injecting this material subcutaneously in mice, optical imaging was used to quantitatively monitor phagocytosis and dissolution of fibrils concurrently. Histologic evaluation of the residual fibril masses revealed the presence of CD68 + , F4/80 + , ionized calcium binding adaptor molecule 1 - macrophages containing Congo red-stained fibrils as well as neutrophil-associated proteins with no evidence of intact neutrophils. These data suggest an early infiltration of neutrophils, followed by extensive phagocytosis of the light chain fibrils by macrophages, leading to dissolution of the mass. Optical imaging of this novel murine model, coupled with histologic evaluation, can be used to study the cellular mechanisms underlying dissolution of synthetic amyloid-like fibrils and human amyloid extracts. In addition, it may serve as a test bed to evaluate investigational opsonizing agents that might serve as therapeutic agents for light chain-associated amyloidosis., (Copyright © 2019 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

Staphylococcus aureus is the causative agent of multiple infections, including bacteremia, infective endocarditis, osteomyelitis, septic arthritis, and prosthetic device infections. We report here the first whole-genome sequence for four S. aureus sequence type 398 isolates from clinical cases of osteomyelitis in four goats with a history of orthopedic surgery.

Bacterial infections from Staphylococcus pseudintermedius are the most common cause of skin infections (pyoderma) affecting dogs. Two component pore-forming leukocidins are a family of potent toxins secreted by staphylococci and consist of S (slow) and F (fast) components. They impair the innate immune system, the first line of defense against these pathogens. Seven different leukocidins have been characterized in Staphylococcus aureus, some of which are host and cell specific. Through genome sequencing and analysis of the S. pseudintermedius secretome using liquid chromatography mass spectrometry we identified two proteins, named "LukS-I" and "LukF-I", encoded on a degenerate prophage contained in the genome of S. pseudintermedius isolates. Phylogenetic analysis of LukS-I components in comparison to the rest of the leukocidin family showed that LukS-I was most closely related to S. intermedius LukS-I, S. aureus LukE and LukP, whereas LukF-I was most similar to S. intermedius LukF-I S. aureus gamma hemolysin subunit B. The killing effect of recombinant S. pseudintermedius LukS-I and LukF-I on canine polymorphonuclear leukocytes was determined using a flow cytometry cell permeability assay. The cytotoxic effect occurred only when the two recombinant proteins were combined. Engineered mutant versions of the two-component pore-forming leukocidins, produced through amino acids substitutions at selected points, were not cytotoxic. Anti-Luk-I produced in dogs against attenuated proteins reduced the cytotoxic effect of native canine leukotoxin which highlights the importance of Luk-I as a promising component in a vaccine against canine S. pseudintermedius infections., Competing Interests: We have the following interests. We have a patent application for PCT/US18/46281 "Staphylococcus pseudintermedius virulence factor compositions." This non-provisional application was filed by the University of Tennessee and is not associated with any transfer or sale of rights or ownership or any revenue. There are no further patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

Coagulase activation of prothrombin by staphylococcus induces the formation of fibrin deposition that facilitates the establishment of infection by Staphylococcus species. Coagulase activity is a key characteristic of Staphylococcus pseudintermedius ; however, no coagulase gene or associated protein has been studied to characterize this activity. We report a recombinant protein sharing 40% similarity to Staphylococcus aureus coagulase produced from a putative S. pseudintermedius coagulase gene. Prothrombin activation by the protein was measured with a chromogenic assay using thrombin tripeptide substrate. Stronger interaction with bovine prothrombin than with human prothrombin was observed. The S. pseudintermedius coagulase protein also bound complement C3 and immunoglobulin. Recombinant coagulase facilitated the escape of S. pseudintermedius from phagocytosis, presumably by forming a bridge between opsonizing antibody, complement, and fibrinogen. Evidence from this work suggests that S. pseudintermedius coagulase has multifunctional properties that contribute to immune evasion that likely plays an important role in virulence., (Copyright © 2018 American Society for Microbiology.)

We report here the first whole-genome sequences for 3 strains of Staphylococcus pseudintermedius (112N, 113N, and 114N) isolated in Africa. Samples of this opportunistic pathogen were collected from nasal swabs obtained from healthy carrier dogs in Botswana. The sequence information will facilitate spatial phylogenetic comparisons of staphylococcal species and other bacteria at the genome level., (Copyright © 2018 Abouelkhair et al.)

Background: Staphylococcus pseudintermedius is an opportunistic pathogen that is the major cause of pyoderma affecting dogs. Conventional antimicrobial treatment for infections caused by this organism have failed in recent years due to widespread resistance and alternative treatment strategies are a high priority. Protein A encoded in Staphylococcus aureus by spa protects the bacterium by binding IgG and acts as a superantigen. Staphylococcus pseudintermedius possess two genes orthologous to S. aureus spa, spsP, and spsQ., Methods: SpsQ and SpsQ-M, a non-toxigenic SpsQ, were cloned and expressed as recombinant proteins and their cytotoxic effect on canine B cells was measured. The neutralizing ability of antibody raised against them in clinically healthy dogs was evaluated., Results: S. pseudintermedius SpsQ induced apoptosis of canine B cells. Specific amino acid substitutions diminished SpsQ-M binding to immunoglobulin and its super-antigenic activity, while its antigenicity was maintained. This recombinant, non-toxigenic S. pseudintermedius SpsQ stimulated the production of antibodies in dogs that specifically reacted with SpsQ and greatly diminished its cytotoxic effect on canine B cells., Conclusions: The production of neutralizing antibody suggests that attenuated, non-toxic SpsQ produced in this study is a good candidate for inclusion in a vaccine for use in the treatment and prevention of S. pseudintermedius infections., Abbreviations: SpA: Staphylococcus aureus protein A; SpsP: Staphylococcus pseudintermedius protein A; SpsQ: Staphylococcus pseudintermedius protein A; SpsQ-M: attenuated Staphylococcus pseudintermedius protein A; MRSP: methicillin resistant Staphylococcus pseudintermedius; IgA: immunoglobulin A; IgG: immunoglobulin G; IgM: immunoglobulin M; VH: variable region of immunoglobulin heavy chain; IgBD: immunoglobulin binding domains; MFI: mean fluorescent intensity; SEM: standard error of the mean; PBMC: Peripheral blood mononuclear cells; CD21: complement receptor type 2; ST: Sequence type; OD: Optical density; ORF: open reading frame; PBS: Phosphate buffered saline; Tween 20: Polyethylene glycol sorbitan monolaurate 20; HRP: horseradish peroxidase; TMB- 3,3',5,5'-Tetramethylbenzidine.

Staphylococcus pseudintermedius is an opportunistic pathogen in dogs and the most frequent cause of canine pyoderma. Protein A, a potent virulence factor in S. aureus is encoded by the spa gene. S. pseudintermedius possesses genes seemingly analogous to spa, but the expression and the characteristics of their products have not been directly determined. The purpose of this study was to test isolates from major clonal groups for the presence of spa gene orthologs, quantitate their expression levels, and to characterize protein A in S. pseudintermedius.  From the data, it was observed that S. pseudintermedius isolates express genes analogous to spa in S. aureus. Isolates representing major clonal populations in the United States and Europe, ST68 and ST71 respectively, bound significantly higher amounts of canine IgG than isolates with other genetic backgrounds, suggesting that these isolates have a higher density of protein A on their surface. Also, canine IgG bound to protein A on S. pseudintermedius via its Fc region similar to protein A from S. aureus. The mRNA profile differed based on the bacterial sequence types and correlated to the density of protein A on the bacterial surface. Protein A was also found to be secreted during the exponential growth phase. Phagocytosis experiments with S. pseudintermedius show that blocking of protein A enhanced phagocytosis in whole blood, neutrophils and in DH82 canine macrophage-like cell line. Taken together, the results demonstrate that S. pseudintermedius produces protein A that shares S. aureus protein A's ability to bind the Fc region of immunoglobulins and may serve as a potential virulence factor by evading the host immune system.

Protein A in Staphylococcus aureus is encoded by the spa (staphylococcal protein A) gene and binds to immunoglobulin (Ig). The S. aureus strain Wood 46 has been variously reported as protein A-deficient and/or spa negative and used as a control in animal models of staphylococcal infections. The results of this study indicate that Wood 46 has normal spa expression but transcribes very low levels of the srtA gene which encodes the sortase A (SrtA) enzyme. This is consistent with unique mutations in the srtA promoter. In this study, a low level of sortase A explains deficient anchoring of proteins with an LPXTG motif, such as protein A, fibrinogen-binding protein and fibronectin-binding proteins A and B on to the peptidoglycan cell wall. The activity of secreted protein A is an important consideration for use of Wood 46 in functional experiments and animal models.

Whole-genome sequencing of Staphylococcus xylosus strain JW2311 from bovine mastitis milk identified the novel 49.3-kb macrolide-lincosamide-streptogramin B (MLS B ) resistance plasmid pJW2311. It contained the macrolide resistance gene mph (C), the macrolide-streptogramin B resistance gene msr (A), and the new MLS B resistance gene erm (48) and could be transformed into Staphylococcus aureus by electroporation. Functionality of erm (48) was demonstrated by cloning and expression in S. aureus ., (Copyright © 2017 American Society for Microbiology.)

Here, we report the first complete genome sequence of the Staphylococcus aureus strain Wood 46. Wood 46 has played an important role in understanding the virulence and pathogenesis of S. aureus infections. This report will assist efforts in vaccine development against methicillin-resistant S. aureus (MRSA) infections., (Copyright © 2017 Balachandran et al.)

Clinical reference textbooks lack data for pyrrolidonyl arylamidase (PYR) activity in Staphylococcus delphini This study evaluated PYR activities of 21 S. delphini strains by reference broth, rapid disc, and rapid slide methods. Species and subgroup identifications were confirmed by nucleic acid-based methods and included nine group A and 12 group B strains. Testing by rapid PYR methods with products from four manufacturers was performed at two testing locations, and, with the exception of one strain tested at one location using reagents from one manufacturer, each S. delphini strain tested positive for PYR activity. Therefore, PYR may be a useful single-test adjunct for distinguishing Staphylococcus aureus from S. delphini and other members of the Staphylococcus intermedius group., (Copyright © 2017 American Society for Microbiology.)

We report the first complete genome sequence of LMG 22219 (=ON 86 T = CCUG 49543 T ), the Staphylococcus pseudintermedius type strain isolated from feline lung tissue. This sequence information will facilitate phylogenetic comparisons of staphylococcal species and other bacteria at the genome level., (Copyright © 2017 Abouelkhair et al.)

Background: Corns are hard protuberances that occur on the digital footpads of Greyhound dogs. The cause of these lesions is unknown and there is little information about them in the veterinary literature. We received anecdotal examples of dog to dog spread of corns suggesting an infectious cause. The aim of this study was to determine if papillomavirus (PV) is associated with Greyhound corns., Methods: We examined four corns from two unrelated adult Greyhound dogs that resided in Florida and Washington, respectively, for PV by PCR. The samples were obtained by owner coring of two lesions from one dog and laser removal of two lesions from the other dog. Total nucleic acid was extracted and DNA was amplified using two PCR primer sets that have been shown to amplify a broad range of PVs from humans and animals: FAP59/ FAP64 and MY11/ MY09. The DNA sequences were compared with all sequences in GenBank. Formalin-fixed, paraffin-embedded tissue from the footpads of four dogs with other inflammatory dermatoses were also examined., Results: PV DNA was amplified from all four corn lesions, while no PV DNA was amplified from other tissues. Comparison of the 444-bp sequences amplified by the MY11/ MY09 primers identified two different PVs. One showed 96% nucleotide sequence similarity with the L1 gene of canine PV type 12. The other showed 78% similarity to canine PV type 16 and, therefore, represents a novel PV. In one of the corns, infection by two of the identified PVs was found., Discussion: These results suggest PV infection could be involved in the pathogenesis of corns in Greyhound dogs., Competing Interests: The authors declare there are no competing interests.

An organism reported in the early literature to be a rare cause of cervical lymphadenitis in guinea pigs, Streptobacillus moniliformis, has been reclassified as Caviibacter abscessus We describe a series of sequential cases of abscesses in guinea pigs that were presented to our clinic from which the only agent isolated was a unique, serum-requiring bacterium. Discrete colonies were not detected in 6.5% CO 2 or anaerobically on routine primary isolation media containing up to 5% whole sheep blood, with and without cysteine, vitamin K, and hemin supplementation after 7 days of incubation at 37°C. Based on subsequently determined growth requirements, the organisms were best described as serum-requiring, aerotolerant anaerobes. Colonies were detectable within 24 h at 37°C in an anaerobic atmosphere on a mycoplasma agar-based medium containing 10% pig serum and reached 3 mm in diameter within 3-5 days. Microscopic appearance consisted of small gram-negative rods and coccobacilli with occasional filaments. However, in direct smears from clinical specimens and from weak or dysgonic growth on plates incubated under suboptimal growth conditions (e.g., in 6.5% CO 2 ), irregular rods with occasional small bulbous forms or numerous long wavy filaments were observed. All of the isolates generated unique spectral profiles similar to that of C. abscessus when examined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Phylogenetic analysis of partial 16S rRNA gene sequences showed that the isolates were identical to each other and shared 99.9% sequence identity with C. abscessus., (© 2016 The Author(s).)

We report the first complete genome sequences of three predominant clones (ST68, ST71, and ST84) of methicillin-resistant Staphylococcus pseudintermedius in North America. All strains were isolated from canine infections and have different SCCmec elements and antibiotic resistance gene patterns., (Copyright © 2016 Riley et al.)

Feline immunodeficiency virus (FIV) is an important infectious agent of cats. Clinical syndromes resulting from FIV infection include immunodeficiency, opportunistic infections, and neoplasia. In our study, a 5' long terminal repeat/gag region-based reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) was developed to amplify all known FIV strains to facilitate point-of-need FIV diagnosis. The RT-iiPCR method was applied in a point-of-need PCR detection platform--a field-deployable device capable of generating automatically interpreted RT-iiPCR results from nucleic acids within 1 hr. Limit of detection 95% of FIV RT-iiPCR was calculated to be 95 copies standard in vitro transcription RNA per reaction. Endpoint dilution studies with serial dilutions of an ATCC FIV type strain showed that the sensitivity of lyophilized FIV RT-iiPCR reagent was comparable to that of a reference nested PCR. The established reaction did not amplify any nontargeted feline pathogens, including Felid herpesvirus 1, feline coronavirus, Feline calicivirus, Feline leukemia virus, Mycoplasma haemofelis, and Chlamydophila felis. Based on analysis of 76 clinical samples (including blood and bone marrow) with the FIV RT-iiPCR, test sensitivity was 97.78% (44/45), specificity was 100.00% (31/31), and agreement was 98.65% (75/76), determined against a reference nested-PCR assay. A kappa value of 0.97 indicated excellent correlation between these 2 methods. The lyophilized FIV RT-iiPCR reagent, deployed on a user-friendly portable device, has potential utility for rapid and easy point-of-need detection of FIV in cats., (© 2015 The Author(s).)

Objective: To evaluate the ability of small interfering RNAs (siRNAs) to inhibit in vitro viral replication and gene expression of feline coronavirus (FCoV)., Sample: Cell cultures of Crandell-Rees feline kidney cells., Procedures: 5 synthetic siRNAs that each targeted a different region of the FCoV genome were tested individually and in various combinations for their antiviral effects against 2 strains of FCoV (feline infectious peritonitis virus WSU 79-1146 and feline enteric coronavirus WSU 79-1683) in cell cultures. Tested combinations targeted the FCoV leader and 3' untranslated region, FCoV leader region and nucleocapsid gene, and FCoV leader region, 3' untranslated region, and nucleocapsid gene. For each test condition, assessments included relative quantification of the inhibition of intracellular viral genomic RNA synthesis by means of real-time, reverse-transcription PCR analysis; flow cytometric evaluation of the reduction of viral protein expression in infected cells; and assessment of virus replication inhibition via titration of extracellular virus with a TCID₅₀ infectivity assay., Results: The 5 siRNAs had variable inhibitory effects on FCoV when used singly. Combinations of siRNAs that targeted different regions of the viral genome resulted in more effective viral inhibition than did individual siRNAs that targeted a single gene. The tested siRNA combinations resulted in approximately 95% reduction in viral replication (based on virus titration results), compared with findings in negative control, nontargeting siRNA-treated, FCoV-infected cells., Conclusions and Clinical Relevance: In vitro replication of FCoV was specifically inhibited by siRNAs that targeted coding and noncoding regions of the viral genome, suggesting a potential therapeutic application of RNA interference in treatment of feline infectious peritonitis.

Nonhemolytic isolates of catalase-positive, Gram-positive cocci were observed that produced strong synergistic hemolysis with colonies of Staphylococcus pseudintermedius and Staphylococcus schleiferi subsp. coagulans in primary cultures of clinical specimens from dogs. The isolates appeared similar to nonhemolytic staphylococci but were not identified with conventional biochemical tests or commercial biochemical test kits. Partial 16S ribosomal RNA gene sequences from each of 9 tested isolates were nearly identical to that of Rothia nasimurium. The isolates produced positive Christie-Atkins-Munch-Peterson test reactions with Staphylococcus aureus (ATCC strain 25923) and Staphylococcus intermedius Hajek (ATCC strain 49052). The latter was identified in the current study as S. pseudintermedius. Rothia nasimurium appears to be a previously unreported commensal organism in dogs. Its potential significance as an opportunistic contributor to polymicrobial infections in dogs is not known., (© 2014 The Author(s).)

Squamous cell carcinoma (SCC) is the most common tumor arising in the periocular and penile areas of horses. Both ultraviolet radiation and papillomaviruses have been implicated in the pathogenesis of SCC in various species, including the horse. This retrospective study used polymerase chain reaction (PCR) to detect papillomavirus DNA in archival biopsy samples from equine periocular and penile SCC from 3 different geographic areas (northeast, southeast, and central United States). Forty-two periocular SCCs were tested; none contained papillomavirus DNA. Twenty-two penile SCCs were tested, and papillomavirus DNA was identified in 10 (43%) cases. Sequencing of the PCR products revealed homology with Equus caballus papillomavirus 2 (EcPV-2). No geographic distribution in the detection of papillomavirus was identified. Penile SCCs were significantly more likely to be papillomavirus positive than the periocular SCCs (P < 0.001). The role of papillomavirus in the development of penile SCC requires further investigation. The differing pathogeneses of periocular and penile SCC suggest that the tumors may respond differently to treatment.

We report the first genome sequence of the methicillin-resistant Staphylococcus pseudintermedius (MRSP) strain E140, isolated from a canine bite wound infection in Denmark. This strain represents the dominant clonal lineage associated with canine MRSP infections in Europe.

Objective: To evaluate agents used for delivery of small interfering RNAs (siRNAs) into feline corneal cells, toxicity of the delivery agents, and functionality of anti-feline herpesvirus 1 (FHV-1)-specific siRNA combinations., Sample: Feline primary corneal cells and 19 six-month-old colony-bred cats., Procedures: siRNA delivery into corneal cells via various delivery agents was evaluated via flow cytometric detection of labeled siRNAs. Cellular toxicity was evaluated with a proliferation assay. Functionality was tested via quantitative reverse transcriptase PCR assay, plaque assay, and flow cytometry. In vivo safety was evaluated with an ocular scoring method following topical application of delivery agents containing siRNAs into eyes. Corneal biopsy specimens were used to assess safety and uptake of siRNAs into corneal cells., Results: Use of 3 delivery agents resulted in > 95% transfection of primary corneal cells. Use of a peptide for ocular delivery yielded approximately 82% transfection of cells in vitro. In cultured corneal cells, use of the siRNA combinations resulted in approximately 76% to 89% reduction in FHV-1-specific mRNA, 63% to 67% reduction of FHV-1-specific proteins in treated cells, and 97% to 98% reduction in FHV-1 replication. The agents were nonirritating in eyes, caused no substantial clinical ocular signs, and were nontoxic. Histologically, corneal epithelium and stroma were normal in treated cats. However, none of the agents were effective in delivering siRNAs into the corneal cells in vivo., Conclusions and Clinical Relevance: The tested anti-FHV-1-specific siRNAs could potentially be used as a treatment for FHV-1 if a successful means of in vivo delivery can be achieved.

Staphylococcus pseudintermedius is an opportunistic pathogen in dogs. Four housekeeping genes with allelic polymorphisms were identified and used to develop an expanded multilocus sequence typing (MLST) scheme. The new seven-locus technique shows S. pseudintermedius to have greater genetic diversity than previous methods and discriminates more isolates based upon host origin.

Cefoxitin disk diffusion susceptibility testing is a recommended screening method for the detection of methicillin resistance in human isolates of Staphylococcus aureus and coagulase-negative staphylococci. A retrospective analysis of 1,146 clinical isolates of Staphylococcus pseudintermedius from dogs was conducted to determine if screening by the cefoxitin disk method can be similarly useful with S. pseudintermedius. The distribution of cefoxitin growth inhibition zone diameters within this collection was bimodal and correlated well with the results of methicillin resistance gene (mecA) detection by polymerase chain reaction. Of the isolates, 5% had discordant results and, when retested, 84% of these were in agreement. While a greater diversity of isolates and interlaboratory comparisons must be tested, the current study suggests that an epidemiological breakpoint (of approximately ≤ 30 mm = resistant; ≥ 31 = susceptible) can be established to predict methicillin resistance in S. pseudintermedius.

Background: This study evaluated the exposure of dogs to three different Ehrlichia spp. in the south and central regions of the United States where vector-borne disease prevalence has been previously difficult to ascertain, particularly beyond the metropolitan areas., Methods: Dog blood samples (n = 8,662) were submitted from 14 veterinary colleges, 6 private veterinary practices and 4 diagnostic laboratories across this region. Samples were tested for E. canis, E. chaffeensis and E. ewingii specific antibodies using peptide microtiter ELISAs., Results: Overall, E. canis, E. chaffeensis and E. ewingii seroprevalence was 0.8%, 2.8%, and 5.1%, respectively. The highest E. canis seroprevalence (2.3%) was found in a region encompassing Arkansas, Louisiana, Oklahoma, Tennessee and Texas. E. chaffeensis seroreactivity was 6.6% in the central region (Arkansas, Kansas, Missouri, and Oklahoma) and 4.6% in the southeast region (Georgia, Maryland, North Carolina, South Carolina, Tennessee and Virginia). Seroreactivity to E. ewingii was also highest in the central region (14.6%) followed by the southeast region (5.9%). The geospatial pattern derived from E. chaffeensis and E. ewingii seropositive samples was similar to previous reports based on E. chaffeensis seroreactivity in white-tailed deer and the distribution of human monocytic ehrlichiosis (HME) cases reported by the CDC., Conclusions: The results of this study provide the first large scale regional documentation of exposure to E. canis, E. chaffeensis and E. ewingii in pet dogs, highlighting regional differences in seroprevalence and providing the basis for heightened awareness of these emerging vector-borne pathogens by veterinarians and public health agencies.

The present case report describes isolation of the fungus Oxyporus corticola from multiple lymphocutaneous tissues of a Beagle dog. Until recently, this fungus had not been reported in the human or veterinary medical literature as a cause of animal disease. A single previous report also involved infection in a German Shepherd Dog, a breed with reported increased susceptibility to disseminated fungal infection and dysfunctional immune response. Isolates were non-sporulating and required molecular identification methods for prompt differentiation from other fungal pathogens. Risk factors for infection with O. corticola are unknown.

Isolates of gram-negative anaerobic bacteria from reptiles have only occasionally been identified to the genus and species level in the veterinary medical literature. In particular, reports identifying Porphyromonas spp. from infections in reptiles are scarce. The present report describes unique Porphyromonas isolates obtained from necrosuppurative infections in central bearded dragons (Pogona vitticeps). The isolates grew in the presence of oxygen, were strongly hemolytic, and did not produce detectable black, iron porphyrin pigment. Biochemical identification kit numeric biocodes gave high but unreliable probabilities (>99.9%) for identification as Porphyromonas gingivalis. Partial 16S ribosomal RNA gene sequences of the isolates were identical to each other and shared 91% identity with those of Porphyromonas gulae. The isolates may represent a new reptile-associated Porphyromonas species.

Objective: To characterize the L1 gene of papillomaviruses detected in epithelial lesions of cats and to determine the relationship between those L1 gene nucleotide sequences and known L1 gene sequences of human and feline papillomaviruses., Sample Population: 10 tissue samples of epithelial lesions from 8 cats., Procedures: DNA was extracted from tissue samples. Primers were designed to amplify the L1 gene of papillomaviruses. Amplicons of DNA were sequenced; nucleotide sequences were compared with known L1 gene nucleotide sequences of papillomaviruses and used for phylogenetic analysis., Results: Tissue samples were obtained from lesions (diagnosed as dysplasia [n=1], squamous cell carcinoma in situ [3], or squamous cell carcinoma [6]) of the skin (9) and oral mucosa [1]. Two amplicons had 99% homology with the L1 gene nucleotide sequence of human papillomavirus type 38b subtype FA125. Another amplicon had 84% homology with the L1 gene nucleotide sequence of human papillomavirus type 80 and was considered to be a new type of papillomavirus. Phylogenetic tree analysis revealed that these 3 papillomaviruses were grouped into 2 clades that were not similar to the clades of Felis domesticus papillomavirus type 1 or F domesticus papillomavirus type 2 (FdPV2). The remaining 7 amplicons had 98% to 100% homology with the L1 gene nucleotide sequence of FdPV2. Phylogenetic tree analysis revealed that those 7 papillomaviruses were grouped nto a single clade with FdPV2., Conclusions and Clinical Relevance: Results support the likelihood of transmission of papillomaviruses between humans and cats.

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3-5 µm in diameter, Gomori methenamine silver-positive structures, consistent with Lycoperdon spores. An approximately 750-base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesting that airway sampling may assist in diagnosing lycoperdonosis.

Objective: To evaluate the ability of 5 small interfering RNAs (siRNAs) targeting mRNA of the feline herpesvirus-1 (FHV-1) DNA polymerase gene to reduce in vitro viral replication and gene expression of FHV-1, to evaluate combinations of these siRNAs with siRNAs that target the glycoprotein D gene of FHV-1, and to determine the combination or combinations of siRNAs that yield the greatest inhibition of in vitro viral replication., Sample Population: Cultured Crandell-Rees feline kidney (CRFK) cells., Procedures: CRFK cells were transfected with siRNAs designed to target mRNA of the FHV-1 DNA polymerase gene. Effective treatment was determined by quantification of the inhibition of mRNA available for DNA polymerase translation, viral protein production, and viral replication. Combinations of 2 siRNAs that target mRNA of the FHV-1 DNA polymerase gene and 2 siRNAs that target the mRNA of the essential FHV-1 glycoprotein D gene were evaluated for the ability to inhibit viral replication., Results: Verified by a reduction in viral gene expression, 2 of the 5 siRNAs designed to target mRNA of the FHV-1 DNA polymerase gene significantly suppressed viral replication. Two combinations of siRNAs that target mRNA of the FHV-1 DNA polymerase gene, the FHV-1 glycoprotein D gene, or both also significantly suppressed viral replication., Conclusions and Clinical Relevance: Combinations of siRNAs that target mRNA of the FHV-1 DNA polymerase gene, FHV-1 glycoprotein D gene, or both could potentially be used as a treatment for the prevention of clinical disease associated with FHV-1 infection.