19th SIAV/ItVAS INTERNATIONAL SEMINAR on VETERINARY ACUPUNCTURE

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1 19th SIAV/ItVAS INTERNATIONAL SEMINAR on VETERINARY ACUPUNCTURE Giuseppina Brocherel Sezione di Arezzo IZSLT CeMIV Arezzo - Italy giuseppina.brocherel@izslt.it Francesco Longo CIAV/ItVAC SIAV/ItVAS Bologna Italy longo.agovet@katamail.com

2 Anemic/Haemolytic Syndromes Theileria equi, Babesia caballi and Anaplasma phagocytophilum (zoonosis), transmitted by ticks of the family Ixodidae are among the causative agents responsible for anemic / hemolytic syndromes in equines Theileria equi e Babesia caballi Equine Piroplasmosis Anaplasma phagocytophilum Equine Anaplasmosis (Equine granulocytic anaplasmosis)

3 Vectors Species Anaplasma Phagocytophilum Ixodes Ricinus Babesia Caballi Rhipicephalus Dermacentor Haemaphysalis Theileri Equi Rhipicephalus Dermacentor Hyalomma

4 Factors Affecting Transmission The movement of these clinically healthy carrier animals into regions with competent tick vectors, where they can be a source for infection of naïve horse populations, is the basis for the spread of these parasites. Increased international movement of horses, development of tick populations with increased resistance to chemical acaricides, and climate and land usage changes are also important factors increasing the risk of the spread of these parasites to regions previously free of infection. Poor management of equine populations, including inadequate veterinary care, poor nutrition, and overwork, may exacerbate the impacts

Equine Piroplasosis Equine Piroplasmosis (EP), an infectious tick-borne disease of the Equidae, is caused by two intra-erythrocytic hemoprotozoans, Theileria equi (formerly Babesia equi) and Babesia

5 Equine Piroplasosis Equine Piroplasmosis (EP), an infectious tick-borne disease of the Equidae, is caused by two intra-erythrocytic hemoprotozoans, Theileria equi (formerly Babesia equi) and Babesia caballi, in most tropical and subtropical areas, and some temperate zones of the world. In Italy, especially in the centralsouthern regions, EP can be considered endemic, and high rates of seropositive animals are reported

equi parasite in the infected erythrocyte varies from spherical, ovoid or Maltese cross

6 Equine Piroplasmosis Theileria equi is a small piroplasm whereas B. caballi is a larger form. The shape of T. equi parasite in the infected erythrocyte varies from spherical, ovoid or Maltese cross shape. The organism may be found either singly, in pairs, or in tetrads. Babesia caballi organisms are pyriform round or oval in shape and commonly seen singly or in pairs.

7 Theileria equi Reservoir: Horse and Donkey T. equi is either transtadially or intrastadially transmitted (infection moves from the tick midgut directly to the salivary glands) Transplacentar transmission: abortion (most commonly in late gestation), stillbirth, or neonatal infection and can occur across placentas that are histologically normal. (Asymptomatic) Persistent infections infect vector, iatrogenic

8 Babesia caballi Reservoir: Tick B. caballi is usually transmitted transovarially because infection moves from the midgut to the ovaries, where it enters the eggs and subsequently the offspring, but intrastadial transmission also occurs. No persistent infection, low level of parasitaemia

Simpthoms Hyperacute Form: Rare form of disease with only clinical observation being moribund or dead animals Acute Form: Most common form of disease cases Fever that usually exceeds 40 C Reduced

9 Simpthoms Hyperacute Form: Rare form of disease with only clinical observation being moribund or dead animals Acute Form: Most common form of disease cases Fever that usually exceeds 40 C Reduced appetite and malaise Elevated respiratory and pulse rates Congestion of mucous membranes Production of a dark red urine Faecal balls that are smaller and drier than normal Affected animals may appear unthrifty Anemic and/or icteric

Subacute Form Similar to acute form but accompanied by weight loss in affected animals and intermittent fever Mucous membranes vary from pale pink to pink, or pale yellow to

10 Subacute Form Similar to acute form but accompanied by weight loss in affected animals and intermittent fever Mucous membranes vary from pale pink to pink, or pale yellow to bright yellow; Petechiae and/or ecchymosis may also be visible on the mucous membranes Simpthoms Normal bowel movements may be slightly depressed and the animals may show signs of mild

Chronic Form Chronic form Chronic cases usually present nonspecific clinical signs such as mild inappetence, poor performance and a drop in body mass.

11 Chronic Form Chronic form Chronic cases usually present nonspecific clinical signs such as mild inappetence, poor performance and a drop in body mass. Mild anemia, subicterus, and enlarged spleen mild to severe muscle atrophy Pasolini, Maria P. et al. Inflammatory Myopathy in Horses With Chronic Piroplasmosis Veterinary Pathology 55.1 (2017): Crossref. Web.

12 Chronic Form Histologic lesions included: muscle fiber atrophy (100% of cases), degenerative changes (13/16, 81%), and perivascular perimysial and endomysial lymphocytic infiltrates (81% of cases) Reverse transcription polymerase chain reaction (RT-PCR) testing of muscles from affected animals revealed a significant increase of interferon-γ, interleukin-12, and tumor necrosis factor-α gene expression (TNF-alfa) compared to healthy controls. Inflammatory myopathy associated with equine piroplasmosis may involve an autoimmune pathogenesis with upregulation of inflammatory cytokines that may cause myofiber atrophy and degeneration.

move. Triggers dysfunction or suppression of host defenses. Horse infected with A.

13 Anaplasma phagocytophilum Adult horses over 4 years of age generally develop characteristic progressive signs of fever, depression, partial anorexia, limb edema, petechiation, icterus, ataxia, and reluctance to move. Triggers dysfunction or suppression of host defenses. Horse infected with A. phagocytophilum showing distal limb edema. Horse infected with A. phagocytophilum showing petechiation of the oral mucosal membranes.

Diagnosis Estimation of the prevalences of Babesia caballi and Theileria equi infections and Anaplasma phagocytophilum in the regions of Lazio and Tuscany.

14 Diagnosis Estimation of the prevalences of Babesia caballi and Theileria equi infections and Anaplasma phagocytophilum in the regions of Lazio and Tuscany. Development of quantitative methods and analysis of different diagnostic methods in use in relation to the health status of infected subjects (Research current financial year 2011); Scientific Manager: Dr. Scicluna Maria Teresa Both for piropalsmosis and for anaplasmosis, the detection of the microorganism in erythrocytes and in the neutrophil granulocytes from peripheral smear or from buffy coat is possible only during the acute phase of the infection and requires experienced operators

Diagnosis Serological Tests: Indirect Immunofluorescence (IFAT) and ELISA (they do not provide indications about the stage of infection, constituting a problem especially in endemic areas, due to

15 Diagnosis Serological Tests: Indirect Immunofluorescence (IFAT) and ELISA (they do not provide indications about the stage of infection, constituting a problem especially in endemic areas, due to possible cross reactivity, to the subjective reading of the operator, to the high cost of the reagents For piroplasmosis there are animals with chronic infection that have positive reactions at the limit of detection PCR: In endemic areas, in the presence of antibodies in the population, PCR can be the direct method most suitable for diagnostic purposes and to define more precisely the actual health status also for the purpose of handling. Anaplasma phagocitophylum

16 Discrete Results Positive POS / PCR Serology The antibody title remains detectable, even after the parasite has been eliminated Parasitaemia below the detection limit of PCR Possible presence of sequence variations in the 18s rrna gene Serological NEG / PCR Serology Early stage of infestation (precedes antibody response) Antigenic variations that reduce the sensitivity of serological tests

Diseases transmitted by vectors such as Anaplasmosis Babesiosis Theileriosis according to the TCM are TCM due to penetration of Xie Qi (Perturbed Energy), carried by biting insects due

17 Diseases transmitted by vectors such as Anaplasmosis Babesiosis Theileriosis according to the TCM are TCM due to penetration of Xie Qi (Perturbed Energy), carried by biting insects due to a chronic deficit of Wei Qi (Defensive Energy) it affects the layer of Xue (Blood) According to the TCM the point that treats the diseases transmitted by biting insects is PC 6 Nei Guan

18 TCM Diseases transmitted by vectors can determine: Blood Alterations - 血液改变 Xuěyè Gǎibiàn Anoemia 貧血 Pínxuě Haemolytic Syndromes Xuě Zhen

19 TCM Diseases transmitted by vectors can determine: Leukopenia 白細胞減少症 Báixìbāo jiǎnshǎo zhèng Consuptive Illnesses - Xu Lao

20 TCM Wei Qi flows primarily under the skin and in the muscles, especially in the Tendon-Muscular meridians. Wei Qi protects the body from attack by exogenous pathogenic factors such as harsh weather conditions, microorganisms, harmful emotions, and evil spiritual forces. There are three Wei Qi fields extending several feet from the body. All energetic forms of the body, including organs, blood vessels, nervous system, etc., can be accessed and treated through these fields. Wei Qi warms, moistens, and aids in nourishing skin and muscles. Wei Qi adjusts the opening and closing of pores; thus, regulating sweating and the body temperature. It is controlled by the Lungs, which regulates its circulation. Defensive Qi has its root in the Lower Burner (Kidney). It is nourished by the Middle Burner (Stomach and Spleen) and is spread outwards by the Upper Burner (Lung). Wei Qi in the daytime circulates in the Exterior, but at night it goes into the Interior to protect the Yin Organs.

21 TCM SCIENTIFIC EVIDENCES ON ACUPOINTS ST 36 Zu San Li Increases Leuenkepfaline Promotes Lymphocyte Transformation Increases splenic NK lymphocytes Increase Interleukin 2 in the abdominal muscles Increase Interferon γ in the abdominal muscles. SP 6 San Yin Jiao Stimulates the production of T lymphocytes LI 11 Qu Chi + ST 36 Zu San Li (in combination) Increase 2-8 times the serum antibodies

22 TCM SCIENTIFIC EVIDENCES ON ACUPUNCTURE Phagocytosis increased by Neutrophils Increased antibody title in vaccinated subjects De Qi Activities mediated by β endorphins Increases circulating lymphocytes for a few hours Increase Ig Increases peripheral lymphocytes

TCM ACUPUNCTURE THERAPY It involves both

Chinese tradition but also Haemo-acupuncture

according to the principles of Traditional

23 TCM ACUPUNCTURE THERAPY It involves both Needle and Moxa application according to Chinese tradition but also Haemo-acupuncture and Laser-acupuncture. The points indicated below should be selected on the basis of data collected according to the principles of Traditional Chinese Diagnosis in order to achieve an effective therapy for the individual horse

24 TCM BABESIOSIS HYPERACUTE PATHOLOGY EMERGENCY TECHNIQUES Wei Jian GV 26 Ren Zhong Qian-jiu Hou-jiu HT 9 Shao Chong SI 1 Shao Ze LR 1 Da Dun TH 15 Tian Liao HAEMO-ACUPUNCTURE Blood Injection BL 17 Ge Shu BL 18 Gan Shu BL 20 Pi Shu Zao Bai Hui

25 TCM BABESIOSIS ACUTE PATHOLOGY YIN DEFICIENCY of KI and LR Pain and weakness in the lumbar region Knee problems - Fever - Dry maw - Thirst Tongue: red - poorly coat Pulse: fine - quick in a short time anoemia and jaundice appear

26 BABESIOSIS ACUTE PATHOLOGY TCM YIN DEFICIENCY of KI and LR ANOEMIA JAUNDICE BL 18 Gan Shu LR 14 Qi Men LR 3 Tai Chong BL 23 Shen Shu GB 25 - Jing Men KI 3 Tai Xi SP 6 San Yin Jiao GV 9 Zhi Yang CV 15 Jiu Wei BL 17 Ge Shu BL 43 Gao Huang GB 39 Xuan Zhong SP 10 Xuě Hai SP 15 Da Heng LU 7 Lie Que CV 17 Shan Zhong BL 19 Dan Shu GB 24 Ri Yue GB 25 Jing Men GB 34 - Yang Ling Quan GB 38 Yang Fu SP 4 Gong Sun PC 6 Nei Guan BL 20 Pi Shu LR 13 Zhang Men CV 12 Zhong Wan GV 8 Jing Suo

27 TCM BABESIOSIS SUBACUTE PATHOLOGY DEFICIENCY of QI and XUE Asthenia Mucosae: pale - sometimes with hemorrhagic petechiae or ecchymosis Short breath Brittle hoof and hair Tongue: pale - thin coat Pulse: fine - weak

28 TCM BABESIOSIS SUBACUTE PATHOLOGY DEFICIENCY of QI and XUE ST 36 Zu San Li BL 18 Gan Shu BL 49 Hun Men LR 14 Qi Men BL 17 Ge Shu GV 8 Jing Suo CV 17 Shan Zhong ENERGY LEVEL EXTRAORDINARY MERIDIANS Combined therapy TH 21 Er Men TH 5 Wei Guan TH 1 Guan Chong GB 41 Zu Lin Qi GB 44 Zu Qiao Yin IMMUNE SUPPORT LI 11 Qu Chi + ST 36 Zu San Li

29 TCM BABESIOSIS CHRONIC PATHOLOGY DEFICIENCY of KI and SP YANG Asthenia - Lassitude Lake of appetite - Intestinal problems - Soft stools Myopathy (in different degrees) - Muscle atrophy Poor performance Tongue: pale whitish coat Pulse: fine - deep

30 BABESIOSIS CHRONIC PATHOLOGY TCM DEFICIENCY of KI and SP YANG MYOPATHY ENERGY SUPPORT HAEMO-ACUPUNCTURE Blood Injection BL 20 Pi Shu LR 13 Zhang Men CV 12 Zhong Wan SP 4 Gong Sun PC 6 Nei Guan BL 23 Shen Shu GB 34 - Yang Ling Quan GB 41 Zu Lin Qi TH 5 Wei Guan BL 18 Gan Shu GV 8 Jing Suo CV 7 Yin Jiao LR 3 Tai Chong GV 9 Zhi Yang CV 15 Jiu Wei BL 17 Ge Shu BL 18 Gan Shu Zao Bai Hui GB 25 - Jing Men Hwato Jiaji L3

31 Conclusions Before starting any therapeutic intervention it is essential to know what kind of pathogen is involved and if they are present at the same time No babesicide drug is able to definitively eliminate Theileria equi The drug accumulates in the liver and kidneys to allow the infected cells to repeatedly come into contact with high concentrations SIDE EFFECTS Stimulation of the immune system is fundamental TCM Conclusions Acupuncture and TCM play different roles: It is a valid therapy to treat babesiosis in all of its phases It supports the immune system It ensures a high level of well-being and energy for affected horses It has no side effects!!

32 19th SIAV/ItVAS INTERNATIONAL SEMINAR on VETERINARY ACUPUNCTURE Giuseppina Brocherel Francesco Longo T H A N K S

DERMATOMES and ACUPUNCTURE in the HORSE

XVI ItVAS INTERNATIONAL SEMINAR X VAM/ECVA MEETING Palermo (I) October 23 24 25, 2015 DERMATOMES and ACUPUNCTURE in the HORSE Francesco LONGO DVM, Repr. Sp. S.I.A.V. It.V.A.S. (Italian Veterinary Acupuncture