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SALCI (H.) AND COLLABORATORS114

Introduction

Keratoconjunctivitis is a multi-factorial ocular disease in farm animals that can result from allergies, viruses (infectious bovine rhinotracheitis etc.), bacteria (Moraxella bovis) and parasites (Thelazia spp.) [2, 4, 12]. Although many systemic diseases in cattle have different types of keratoconjunctivitis and similar ocular symptoms, the well-known primer etiologic agent of infectious keratoconjuctivitis is M bovis, which is contagious and results in suffering and economic loses [4, 8-12].

Pseudomonas stutzeri is an aerobic, non-fermentative, Gram-negative microorganism and usually lives in contaminated areas or in pond water, soil and hospitals [5-7]. This microorganism can be isolated in humans with poor health condition or the elderly [6]. Particularly, if the host immunity is compromised; P stutzeri becomes pathogen [7]. The reported diseases of P stutzeri in humans are pneumonia,

meningitis and ocular problems including endophtalmitis, corneal perforation and ocular discharge [5-7]. Although Pseudomonas stutzeri has been encountered in the ocular discharge of the Bustards, it has not been determined as a primary cause of keratoconjunctivitis in animals until now. Therefore, this clinical study is aimed to report P stutzeri microorganism as a causative agent of infectious keratoconjunctivitis in cattle.

Case History

A sample of 20 Holstein breed cattle with eye problems (n=20) in a herd (consist of 165 cattle) was taken in spring near the city of Bursa within Turkey. The owner of the cattle had experienced M bovis infections amongst his cattle in the past, therefore, had segregated the diseased cattle.

Before any clinical examinations or medical applications were made to the affected eyes, microbiological samples were

SUMMARY

The aim of this clinical case was to report Pseudomonas stutzeri microorganism as a causative agent of infectious keratoconjunctivitis in cattle. In totally, 20 Holstein cattle (n=20) with keratoconjunctivitis in a herd were referred. Initially, microbiological samples were taken from the conjunctival recessus of all cases with sterile swaps for culture and antibiotic susceptibility. Following clinical examinations, the cases were classified as mild, moderate and severe according to type of eye disease. As an initial therapy, local 3% acid boric irrigation was given, perpalpebral oxytetracicline was injected and tetracycline pomade was applied before diagnosis took place. The prevalence of the disease was 12.12% in the herd and there was 9 bilateral (n=9) and 11 unilateral (n=11) eye problems. Epiphora and conjunctivitis were the clinical finding in mild cases (n=17). Photophobia, blepharospasm, chemosis, keratoconjunctivitis and ocular pain were observed in moderate cases (n=2). Anterior uveitis and associated symptoms of moderate cases was inspected in a severe case (n=1). Microbiologically, P stutzeri was identified in all cases and antibiogram result of this germ pointed out the 75% sensitivity to oxytetracycline. Initial therapy was continued as curative treatment to all cases. All cases healed; healing time for mild and moderate cases were mean 8.3±2.9 and 19.5±2.1 days, respectively. The symptoms of the anterior uveitis in the severe case were terminated at 42 days, but there was minimal lens opacity as a sequel.

Keywords: cattle, infectious keratoconjunctivitis, Pseudomonas stutzeri

RÉSUMÉ

Kératoconjonctivite infectieuse due à Pseudomonas stutzeri chez les bovins

Le but de ce cas clinique était de rapporter un cas de kératoconjonctivite infectieuse contagieuse chez les bovins due à Pseudomonas stutzeri. Vingt bovins Holstein d’un même troupeau avec ont été référés avec des signes de kératoconjonctivite. Dans un premier temps, des échantillons microbiologiques ont été stérilement prélevés dans le récessus conjonctival de tous les animaux pour culture et recherche de la sensibilité aux antibiotiques. Après les examens cliniques, les kératoconjonctivites ont été classées comme légères, modérées ou sévères en fonction des lésions. Une thérapie initiale par irrigation locale d’acide borique a 3% a été effectuée, une injection perpalpebrale d’oxytetracycline a été réalisée et de la tétracycline en pommade a été appliquée avant le diagnostic étiologique. La prévalence de la maladie était de 12,12% dans le troupeau avec des problèmes oculaires bilatéraux dans 9 cas et unilatéraux dans 11 cas. Epiphora et conjonctivite étaient les signes cliniques dans les cas bénins (n = 17). Photophobie, blépharospasme, chemosis, kératoconjonctivite et douleur oculaire ont été observés dans les cas modérés (n = 2). Une uvéite antérieure était présente dans un cas grave (n = 1). Microbiologiquement, P stutzeri a été identifié dans tous les cas. Un antibiogramme a montré 75% de sensibilité à l’oxytétracycline ce qui a entrainé la poursuite du traitement initial.

Mots-clés : bovins, kératoconjonctivite infectieuse, Pseudomonas stutzeri

Infectious keratoconjunctivitis due to Pseudomonas stutzeri in cattle

H. SALCI1*, G. GONCAGUL2, U. CANATAN1

1 Department of Surgery, Faculty of Veterinary Medicine, Uludag University, 16059, Bursa, TURKEY2 Mennan Pasinli Vocational High School, Uludag University, 16059, Bursa, TURKEY

*Corresponding author: hsalci@uludag.edu.tr

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INFECTIOUS KERATOCONJUNCTIVITIS IN CATTLE 115

taken from the conjunctival recessus with sterile swabs and immediately taken to the bacteriology laboratory for culture and antimicrobial susceptibility. Clinical examinations (general examinations: pulsation, respiration, temperature, mucosal membrane colour, capillary refilling time and local lymph node enlargement; ophthalmic examinations: palpebral, conjunctival and corneal appearance with direct inspection and ophthalmoscopy) of the cattle revealed that some ocular symptoms such as ephiphora, conjunctivitis, keratitis, chemosis, blepharospasm and anterior uveitis existed.

According to the severity of the clinical symptoms, the recorded results were grouped as mild, moderate and severe, and the cases were evaluated in these groups as reported previously [11]. The eyes in the mild cases were examined easily, a local anaesthetics agent was dropped to the eyes in moderate and severe cases to facilitate the ocular examination, it also relieved the ocular pain and blepharospasm. Post examination, in all cases, the eyes were irrigated with 3% acid boric solution followed by a routine direct inspection and periocular examinations. Ophtalmoscopy was performed to determinate the presence of the abnormalities such as discharge (serous, mucoid etc.), redness, swelling, oedema of the conjunctiva and corneal transparency, vascularization and changes of the ocular contour, anterior chamber abnormalities, iris and pupillary abnormalities, lens opacity, vitreous and retinal pathologies.

As an initial therapeutic approach in all cases, 200 mg oxytetracycline hydrochloride (Terramycin/LA 200mg/ml, Pfizer, Turkey) was intrapalpebral applied as reported previously (13) every 3 days, along with ointment of oxytetracycline hydrochloride with polymyxin B sulfate (Terramycin ophthalmic ointment, Pfizer, Turkey) which

was applied to the conjunctival pouch of the lower eyelid twice daily, until the culture and antimicrobial susceptibility results were completed.

Microbiologically analysis: Initially all swaps were incubated at 37°C for 24 hours in fluid thioglycollate (BBL; 221196) the next day part of the initial culture was transferred into bloody agar (BBL, 297876) and EMB agar (Eosin-Methylenblue-Lactose-Saccharose) (BBL, 221355) and incubated at 37°C for 24 hours. According to colony morphology and Gram colour features, identified colonies were assessed. For the identification of the bacteria, their direct cultures were performed using BBL Crystal (Becton-Dickinson, Sparks, USA) Gram-Positive and Gram-Negative ID system kits and its computer program.

The antibiotic susceptibility was tested by disk diffusion method as described previously [1]. P stutzeri was evaluated according to National Committee for Clinical Laboratory standard methods [3]. Table II shows the planned antibiotic discs and classification of the susceptibility (sensitive, intermediate and resistance).

Results

The prevalence of the infectious keratoconjunctivitis in the presented herd was 12.12%. There was 9 bilateral (n=9) and 11 unilateral (n=11) eye problems in effected cattle. Clinical results of the cases in groups are detailed in Table I, and also summarized as follows:

The mild cases only had serous-mucoid epiphora and/or conjunctivitis (n=17) (Figure 1). Photophobia, blepharospasm, chemosis, keratoconjunctivitis (generalise hazy blue corneal oedema, red to blue vascularization, non-

SUSCEPTIBILITY (%)

Antibiotic agent Sensitive Intermediate ResistanceTulathromycin (30µg) - - 100Oxytetracycline (30µg) 75 25 -Erythromycin (30µg) - - 100Florfenicol (15µg) - - 100Ceftiofur (30µg) - 25 75Amoxicillin clavulanate (30µg) - - 100Amoxicillin (25µg) - - 100Enrofloxacin (5µg) 100 - -Gentamicin (10µg) 85 15 -Cefquinome (30µg) - 100 -Colistin (50µg) 100 - -Marbofloxacin (10µg) 100Trimethoprim-sulfamethoxazole (25µg) 100

Table II: The applied antibiotic discs and susceptibility result of Pseudomonas stutzeri (%).

Revue Méd. Vét., 2016, 167, 5-6, 114-119

SALCI (H.) AND COLLABORATORS116RI

GH

T EY

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ular

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Tabl

e I:

Clin

ical

and

mic

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ical

resu

lts o

f the

righ

t and

left

eyes

of t

he c

ases

in g

roup

s.

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INFECTIOUS KERATOCONJUNCTIVITIS IN CATTLE 117

ulcerative superficial keratitis) and ocular pain was the encountered results in the moderate cases (n=2) (Figure 2). The severe cases (n=1) had anterior uveitis and associated symptoms of moderate cases (Figure 3).

Microbiological results of the samples taken in groups are given in Table I. As P stutzeri was obtained in all samples, it was considered as a causal agent. Thus antibiotic susceptibility was planned on P stutzeri, the results are given with their percentages in Table II. Taking these susceptibility results into consideration, the initial therapeutic approach was continued in all cases as curative treatment until the recovery.

The cases were controlled for recovery daily, and the exact healing time without any ophthalmic complication in mild and moderate cases occurred: mean 8.3±2.9 and 19.5±2.1 days, respectively. The symptoms of the severe case with anterior uveitis were terminated after 42 days, but minimal lens opacity was observed in this case (Figure 4).

Figure 1: Ocular form of a mild case: epiphora in the left eye.

Figure 2: Clinical appearance of the excessive epiphora, blepharospasm and swelling of the superior eyelid (chemosis) in a moderate case.

Figure 3: Evidence of the anterior uveitis and keratoconjunctivitis in the severe case.

Figure 4: Undesirable sequel in the severe case: minimal lens opacity.

Revue Méd. Vét., 2016, 167, 5-6, 114-119

SALCI (H.) AND COLLABORATORS118

Discussion

M bovis is the most frequently isolated microorganism in infectious keratoconjunctivitis cases, which is widespread and very contagious [10-12]. It is reported yearly and seasonal prevalence varies greatly in different geographic regions. The reported isolation rate increases during the spring (21.4%) and summer (29.3%) [2]. Systemic infections, co-infectious bacteria, viral or abiotic can demonstrate the same ocular signs of infectious keratoconjunctivitis due to M bovis [3, 12]. In presented herd, clinical findings of the P stutzeri infection were similar to M bovis, and the eye problems were also encountered in spring. The prevalence of the infectious keratoconjunctivitis due to P stutzeri in cattle was 12.12%. As a result of the laboratory findings, which were obtained in all ocular samples, P stutzeri was considered to be the major causative pathogen as a new bacterium.

The cases of infectious keratoconjunctivitis present different ocular form related to stage of the disease, persistence and progression of the infection [10]. Ocular pain, excessive lachrymation, watery or purulent ocular discharge, blepharospasm, photophobia, cloudiness appearance of the corneal centre due to different keratitis forms and corneal ulceration are common symptoms in infectious ocular diseases [10-12]. As reported here, epiphora and conjunctivitis were more common ocular problems in mild cases; non-ulcerative superficial keratitis was encountered in all moderate cases. A severe case had anterior uveitis and associated findings of moderate cases. It can be concluded that Pseudomonas stutzeri can be frequently responsible for mild degree infectious keratoconjunctivitis in cattle. Similar to M bovis infection [11], the mild ocular problems also involved unilateral or bilateral eyes, and ocular forms in moderate and severe cases were unilateral and also sporadic.

P stutzeri lives in contaminated areas and responsible to some ocular problems [5-7]. It has been reported as a causative agent of delayed-onset endophtalmitis after uncomplicated cataract surgery in humans [5]. Some specific microorganisms can demonstrate the similar clinical findings of M bovis infection [2], and M bovis infection sometimes associates with concurrent infections [10, 12]. As reported in humans [5], we cultured P stutzeri and the other microorganisms from the conjunctival samples.

Treatment protocols against to P stutzeri should be applied based on antibiotic susceptibility results. P stutzeri is sensitive to some antibiotics such as tetracycline, ceftazidime, gentamicin, ofloxacin, ciprofloxacin, ceftizoxime and piperacillin [5, 7]. In the present cases, P stutzeri was cultured in all samples. Moreover, P stutzeri was unique isolate in two mild, a moderate and severe case. Antibiotic susceptibility pointed out that P stutzeri was sensitive to a similar antibiotic reported earlier [5, 7]. Taking the culture and antibiotic susceptibility results into consideration, P stutzeri was diagnosed as the responsible microorganism in

all three types cases and the initial treatment protocol was continued.

A study comparing the intramusculary and intrapalpebral injections of oxytetracycline reports similar outcomes in cattle with infectious keratoconjunctivitis [13]; thus, intrapalpebral oxytetracycline was applied before and after culture and antibiogram results in cattle.

In conclusion, it should be considered that the other microorganisms such as P stutzeri can be responsible for infectious keratoconjunctivitis in cattle.

Acknowledgement

The authors would like to thank Yuksel BUYUKDENIZ (native speaker from England) for grammatically controls.

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